Objective:To investigate the effects of Eriocitrin on the proliferation and migration of Lung adenocarcinoma(LUAD)cells A549 and H1299,and the mechanism of Epithelial-Mesenchymal Transition(EMT).Methods:The effects of...Objective:To investigate the effects of Eriocitrin on the proliferation and migration of Lung adenocarcinoma(LUAD)cells A549 and H1299,and the mechanism of Epithelial-Mesenchymal Transition(EMT).Methods:The effects of different Eriocitrin on the proliferation of LUAD cells A549 and H1299 were examined by CCK8 method.EMT-associated epithelial calmodulin(E-cadherin and N-cadherin),vimentin,ferroptosis-associated protein SLC7A11,GPX4,FTH were detected by Western Blot and expression of mRNA of EMT marker molecules E-cadherin,N-cadherin,Snail were detected by qRT-PCR.Effects of saccharomyces cerevisiae suberin on ferroptosis in LUAD cells as observed by lipid reactive oxygen species(ROS)assay.Results:Eriocitrin could significantly inhibit the proliferative behavior of LUAD cells A549 and H1299 and showed a certain dose-and time-dependence.Compared with the control group,different concentrations of Eriocitrin could significantly reduce the scratch healing rate after 24 and 48 h of action,and the difference was statistically significant(P<0.01).The expression of ROS is increased,EMT-related protein E-cadherin was increased in LUAD cells A549 and H1299 compared with the control group after the intervention with Eriocitrin.N-cadherin and Vimentin expression was decreased.E-cadherin mRNA expression was increased,and N-cadherin,Snail mRNA expression was decreased,expression of ferroptosis-associated protein SLC7A11,GPX4,FTH was decreased,the difference was statistically significant(P<0.05).Conclusion:Eriocitrin may inhibit the proliferation and migration of LUAD cells by regulating the EMT pathway and has potential application in LUAD prevention and adjuvant chemotherapy.展开更多
AIM:To analyze the relationship between clinical features and epithelial mesenchymal transition(EMT) in retinoblastoma(RB),further to investigate whether miR-200c regulates the EMT and migration of RB cells.METHODS: E...AIM:To analyze the relationship between clinical features and epithelial mesenchymal transition(EMT) in retinoblastoma(RB),further to investigate whether miR-200c regulates the EMT and migration of RB cells.METHODS: Expression of EMT-related markers and tumorrelated factors were detected by immuno-histochemistry analysis in RB tissue from 29 cases. Correlations between their expression and clinical characteristics were analyzed. The regulation effects of miR-200c on EMT-related markers,tumor-related factors were observed in m RNA level and protein level by real-time polymerase chain reaction(PCR) and Western blot,respectively,in Y79 and Weri-rb1 cells. Its effects on migration force of these RB cell lines were also detected with Transwell test.RESULTS: Lower expression of E-cadherin was present in the cases with malignant prognosis. Mi R-200c promoted the expression of E-cadherin and decreased the expression of Vimentin and N-cadherin in Y79 and Weri-rb1 cells. Migration force of RB cells could be inhibited by miR-200c.CONCLUSION: EMT might be associated with bad prognosis in RB. Mi R-200c suppresses the migration of retinoblastomatous cells by reverse EMT.展开更多
Triple Negative Breast Cancer (TNBC) is a malignant form of cancer with very high mortality and morbidity. Epithelial to Mesenchymal Transition (EMT) is the most common pathophysiological change observed in cancer cel...Triple Negative Breast Cancer (TNBC) is a malignant form of cancer with very high mortality and morbidity. Epithelial to Mesenchymal Transition (EMT) is the most common pathophysiological change observed in cancer cells of epithelial origin that promotes metastasis, drug resistance and cancer stem cell formation. Since the information regarding differential gene expression in TNBC cells and cell signaling events leading to EMT is limited, this investigation was done by comparing transcriptomic data generated by RNA isolation and sequencing of a EMT model TNBC cell line in comparison to regular TNBC cells. RNA sequencing and Ingenuity Pathway Software Analysis (IPA) of the transcriptomic data revealed several upregulated and downregulated gene expressions along with novel core canonical pathways including Sirtuin signaling, Oxidative Phosphorylation and Mitochondrial dysfunction events involved in EMT changes of the TNBC cells.展开更多
Objective: To examine the effect of brucine on the migration, invasion, adhesion and expressions of epithelial-to-mesenchymal transition(EMT) markers and matrix metalloproteinases(MMPs) in the highly metastatic breast...Objective: To examine the effect of brucine on the migration, invasion, adhesion and expressions of epithelial-to-mesenchymal transition(EMT) markers and matrix metalloproteinases(MMPs) in the highly metastatic breast cancer cell lines MDA-MB-231 and Hs578-T. Methods: MDA-MB-231 and Hs578-T cells were divided to 4 groups: the control group(0.1% DMSO), and 25, 50 and 100 μmol/L brucine groups. The cell viability was determined using a Cell Titer-Glo? luminescent cell viability. The scratch wound healing assay and tanswell migration assay were used to determine the migration ability of these cells treated by different concentrations of brucine. The proliferation rate, invasive potential and adhesive ability were respectively performed by colony formation assay, transwell invasion assay and adhension assay. The protein and m RNA expressions of EMT biomarkers, MMP-2 and MMP-9 were investigated by real-time reverse transcription polymerase chain reaction and Western blot. Results: Compared with the control group, brucine had little effect on cell viability or proliferation(P>0.05), but led to a dose-dependent decrease on migration, invasion, adhension of MDA-MB-231 and Hs578-T cells(P<0.01). Furthermore, brucine increased the protein and m RNA levels of EMT markers such as E-cadherin and β-catenin in MDA-MB-231 and Hs578-T cells, and decreased the protein and m RNA levels of mesenychmal markers such as vimentin and fibronectin, as well as the expressions of MMP-2 and MMP-9(all P<0.01). Conclusion: Brucine inhibited triple negative breast cancer cells metastasis potentially through EMT reversion and MMP-2 and MMP-9 inhibition.展开更多
BACKGROUND Currently,there are many therapeutic methods for lung adenocarcinoma(LUAD),but the 5-year survival rate is still only 15%at later stages.Epithelial–mesenchymal transition(EMT)has been shown to be closely a...BACKGROUND Currently,there are many therapeutic methods for lung adenocarcinoma(LUAD),but the 5-year survival rate is still only 15%at later stages.Epithelial–mesenchymal transition(EMT)has been shown to be closely associated with local dissemination and subsequent metastasis of solid tumors.However,the role of EMT in the occurrence and development of LUAD remains unclear.AIM To further elucidate the value of EMT-related genes in LUAD prognosis.METHODS Univariate,least absolute shrinkage and selection operator,and multivariate Cox regression analyses were applied to establish and validate a new EMT-related gene signature for predicting LUAD prognosis.The risk model was evaluated by Kaplan–Meier survival analysis,principal component analysis,and functional enrichment analysis and was used for nomogram construction.The potential structures of drugs to which LUAD is sensitive were discussed with respect to EMT-related genes in this model.RESULTS Thirty-three differentially expressed genes related to EMT were found to be highly associated with overall survival(OS)by using univariate Cox regression analysis(log2FC≥1,false discovery rate<0.001).A prognostic signature of 7EMT-associated genes was developed to divide patients into two risk groups by high or low risk scores.Kaplan–Meier survival analysis showed that the OS of patients in the high-risk group was significantly poorer than that of patients in the low-risk group(P<0.05).Multivariate Cox regression analysis showed that the risk score was an independent risk factor for OS(HR>1,P<0.05).The results of receiver operator characteristic curve analysis suggested that the 7-gene signature had a perfect ability to predict prognosis(all area under the curves>0.5).CONCLUSION The EMT-associated gene signature classifier could be used as a feasible indicator for predicting OS.展开更多
<strong>Objective:</strong> Upfront docetaxel use for hormone na<span style="white-space:nowrap;">ï</span>ve advanced prostate cancer is reported that it successfully delayed...<strong>Objective:</strong> Upfront docetaxel use for hormone na<span style="white-space:nowrap;">ï</span>ve advanced prostate cancer is reported that it successfully delayed the progression to hormone refractory stage, though the adequate methodology to obtain the maximum effect is unclear. We investigated these issues from our experiences of upfront docetaxel use with LH-RH antagonist for metastatic hormone sensitive prostate cancer, aiming at the prevention of epithelial-mesenchymal transition (EMT) for apoptosis tolerance. <strong>Patients and Methods:</strong> Of 31 stage IV new prostate cancer patients treated with upfront docetaxel and LH-RH antagonist (Degarelix), 25 patients who could be followed more than 12 months (mean 36.2 months) were analyzed. Docetaxel was used two to three courses basically 75 mg/m2 dose initializing two weeks after the induction of first Degarelix. <strong>Results:</strong> The clinical course was divided clearly to two groups according to prostate specific antigen (PSA) values. Of 25 patients, 12 patient’s PSA did not decrease below 0.1 ng/ml within 6 months (group A) and gradually rose afterwards. PSA in another 13 patients (group B) decreased below 0.1 within 6 months and kept below 0.1 during the follow up period. Although statistically not significant, the initial group A’s PSA was higher than group B’s (average 1308 and 353 ng/ml), however, number of metastasis, Gleason sum, and bone metastatic extent of disease showed no difference between them. Among group B patients, 7 cases had only upfront docetaxel and hormonal therapy, and some of these patients showed only atrophic gland and fibrotic tissue at second prostate biopsy (specimens after more than two years of therapy), suggesting complete response. <strong>Conclusion:</strong> Our study suggested that PSA value at 6 months may predict the outcome of whole therapy. Patients showing PSA less than 0.1 ng/ml at 6 months and requiring no therapy other than docetaxel and hormone may be induced to complete response. Upfront docetaxel with LH-RH antagonist may prevent EMT for obtaining apoptosis tolerance, in case the patient does not have the castration-resistant clone at the beginning of the therapy (group B).展开更多
AIM:To explore the effects of hepatocyte growth factor(HGF)on retinal pigment epithelium(RPE)cell behaviors.METHODS:The human adult retinal pigment epithelial cell line-19(ARPE-19)were treated by HGF or mesenchymalepi...AIM:To explore the effects of hepatocyte growth factor(HGF)on retinal pigment epithelium(RPE)cell behaviors.METHODS:The human adult retinal pigment epithelial cell line-19(ARPE-19)were treated by HGF or mesenchymalepithelial transition factor(MET)inhibitor SU11274 in vitro.Cell viability was detected by a Cell Counting Kit-8 assay.Cell proliferation and motility was detected by a bromodeoxyuridine incorporation assay and a wound healing assay,respectively.The expression levels of MET,phosphorylated MET,protein kinase B(AKT),and phosphorylated AKT proteins were determined by Western blot assay.The MET and phosphorylated MET proteins were also determined by immunofluorescence assay.RESULTS:HGF increased ARPE-19 cells’viability,proliferation and migration,and induced an increase of phosphorylated MET and phosphorylated AKT proteins.SU11274 significantly reduced cell viability,proliferation,and migration and decreased the expression of MET and AKT proteins.SU11274 suppressed HGF-induced increase of viability,proliferation,and migration in ARPE-19 cells.Additionally,SU11274 also blocked HGF-induced phosphorylation of MET and AKT proteins.CONCLUSION:HGF enhances cellular viability,proliferation,and migration in RPE cells through the MET/AKT signaling pathway,whereas this enhancement is suppressed by the MET inhibitor SU11274.HGF-induced MET/AKT signaling might be a vital contributor of RPE cells survival.展开更多
Background:Keloid scarring is a fibroproliferative disease caused by aberrant genetic activation with an unclear underlying mechanism.Genetic predisposition,aberrant cellular responses to environmental factors,increas...Background:Keloid scarring is a fibroproliferative disease caused by aberrant genetic activation with an unclear underlying mechanism.Genetic predisposition,aberrant cellular responses to environmental factors,increased inflammatory cytokines and epithelial-mesenchymal transition(EMT)phenomena are known as major contributors.In this study,we aimed to identify the molecular drivers that initiate keloid pathogenesis.Methods:Bulk tissue RNA sequencing analyses of keloid and normal tissues along with ex vivo and in vitro tests were performed to identify the contributing genes to keloid pathogenesis.An animal model of inflammatory keloid scarring was reproduced by replication of a skin fibrosis model with intradermal bleomycin injection in C57BL/6 mice.Results:Gene set enrichment analysis revealed upregulation of Wnt family member 5A(WNT5A)expression and genes associated with EMT in keloid tissues.Consistently,human keloid tissues and the bleomycin-induced skin fibrosis animal model showed significantly increased expression ofWNT5A and EMT markers.Increased activation of the interleukin(IL)-6/Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway and subsequent elevation of EMT markerswas also observed in keratinocytes co-cultured withWNT5A-activated fibroblasts or keloid fibroblasts.Furthermore,WNT5A silencing and the blockage of IL-6 secretion via neutralizing IL-6 antibody reversed hyperactivation of the STAT pathway and EMT markers in keratinocytes.Lastly,STAT3 silencing significantly reduced the EMT-like phenotypes in both keratinocytes and IL-6-stimulated keratinocytes.Conclusions:Intercellular communication via the WNT5A and STAT pathways possibly underlies a partial mechanism of EMT-like phenomena in keloid pathogenesis.IL-6 secreted from WNT5A-activated fibroblasts or keloid fibroblasts activates the JAK/STAT signaling pathway in adjacent keratinocytes which in turn express EMT markers.A better understanding of keloid development and the role of WNT5A in EMT will promote the development of next-generation targeted treatments for keloid scars.展开更多
SOX4 is highly expressed in gastric cancer(GC)and is associated with tumor grade,metastasis and prognosis,however the mechanism is not clear.We report herein that SOX4 was upregulated and overexpression of SOX4 was as...SOX4 is highly expressed in gastric cancer(GC)and is associated with tumor grade,metastasis and prognosis,however the mechanism is not clear.We report herein that SOX4 was upregulated and overexpression of SOX4 was associated with increased expression of the markers of Epithelialemesenchymal transition(EMT)and stemness in clinic patient samples.In vitro,overexpression of SOX4 promoted the invasion as showed by Transwell assay and stemness of GC cells as assessed by sphere formation assay,which was suppressed by silencing SOX4 with shRNA.Further studies showed that SOX4 up-regulated the expression of EMT transcription factors Twist1,snail1 and zeb1 and stemness transcription factors SOX2 and OCT4,and promoted the nuclear translocation of β-catenin.Moreover,we revealed that TGF-β treatment significantly up-regulated the expression of SOX4 and silencing SOX4 reversed TGF-β induced invasion and sphere formation ability of GC cells.Finally,we showed that SOX4 promoted the lung metastasis and tumor formation ability of gastric cancer cells in nude mice.Our results suggest that SOX4 is a target TGF-β signaling and mediates TGF-β-induced EMT and stem cell characteristics of GC cells,revealing a novel role of TGF-β/SOX4 axis in the regulation of malignant behavior of GC.展开更多
Objective: To explore the effect of connexin 32 (Cx32) on the cell proliferation, migration, invasion of human hepatocellular carcinoma (HCC) cell line Huh7 and its mechanism. Methods: Firstly, bioinformatics techniqu...Objective: To explore the effect of connexin 32 (Cx32) on the cell proliferation, migration, invasion of human hepatocellular carcinoma (HCC) cell line Huh7 and its mechanism. Methods: Firstly, bioinformatics techniques were used to analyze the difference in expression of Cx32 between HCC tissues and normal liver tissues, and the relationship between Cx32 expression and important clinicopathological features of HCC was also explored. Subsequently, Cx32 expression in HCC cell lines and normal hepatic epithelial cell line was detected in vitro. Huh7 cell line with stable over⁃expression of Cx32 was further established, and the change in cell proliferation ability was measured by MTT assay, changes in migration and invasion capacities were detected by wound⁃healing assay and transwell assay, on this cell line. Finally, western blot and immunofluorescence (IF) were used to investigate the alterations of expression of epithelial⁃mesenchymal transition (EMT) markers. Results: Bioinformatics analyses showed that Cx32 mRNA and protein expression levels in HCC tissues were lower than those in normal liver tissues, and the mRNA expression level of Cx32 was negatively correlated with T stage, histological grade and clinical stage of HCC patients (all P<0.05). Results of in vitro experiments revealed Cx32 protein expression in different HCC cell lines was down⁃regulated compared to that in normal hepatic epithelial cell line LO2. Cx32 stably over⁃expressed (Cx32 OE) Huh7 cell line was successfully constructed by lentivirus infection and showed high expression of Cx32 protein in the cell line. Compared to the control group and (or) the negative control (NC) group, the Cx32 OE group exhibited decreased OD490 value, wound healing rate and invasive cell number (all P<0.05). Furthermore, an increase in the expression of epithelial marker E⁃cadherin, and a decrease in the expression of mesenchymal markers Snail and Vimentin, were observed in Cx32⁃OE Huh7 cell line. Conclusion: Cx32 is low expressed in HCC tissues and cells, while the proliferation, migration and invasion ability of Huh7 cells can be inhibited by over⁃expression of Cx32, of which the underlying mechanism may be related to the inhibition of EMT process.展开更多
Background:Epithelial to mesenchymal transition(EMT)is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma(HCC).Follistatin(FST)family members are considered to ...Background:Epithelial to mesenchymal transition(EMT)is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma(HCC).Follistatin(FST)family members are considered to be an attractive therapeutic targets and prognostic indicators in cancers.As a derivative of FST,Follistatin Like 5(FSTL5)may play a similar role in HCC cells.This study aimed to investigate the expression and function of FSTL5 in HCC and its role in EMT.Methods:FSTL5,E-cadherin and vimentin in HCC,and paracancerous tissues were detected by immunohistochemistry.Correlation of FSTL5 expression with overall survival was assessed.The proliferation and invasion of HCC cell lines SK-Hep1 and MHCC-LM3 were analyzed by cell counting kit-8 and Transwell assays.The expression of FSTL5,E-cadherin,and vimentin in HCC cells was examined by polymerase chain reaction and Western blot analysis.T-test was used to analyze the difference in proliferation and invasion ability between groups.The Spearman rank correlation test was used to detect the correlation between the expression of FSTL5 and E-cadherin or vimentin.Results:The expression of FSTL5 in HCC was lower than that in paracancerous tissues(9.97%vs.82.55%,χ2=340.15,P<0.001).Patients with high FSTL5 expression had a better prognosis(χ2=8.22,P=0.004)and smaller tumor diameter(χ2=45.52,P<0.001),less lymph node metastasis(χ2=5.58,P=0.02),earlier tumor node metastasis stage(χ2=11.29,P=0.001),a reduced number of tumors(χ2=5.05,P=0.02),lower alpha-fetoprotein value(χ2=24.36,P<0.001),more probability of hepatitis carrying(χ2=40.9,P<0.001),and better liver function grade(χ2=5.21,P=0.02).Immunohistochemistry showed that FSTL5 expression in HCC tissues was positively correlated with E-cadherin expression(r=0.38,P<0.001)and negatively correlated with vimentin expression(r=-0.385,P<0.001).Furthermore,over-expression of FSTL5 up-regulated the expression of E-cadherin and down-regulated the expression of vimentin in SK-Hep1(negative control[NC]vs.FSTL5-interfering group[Lv-FSTL5]:E-cadherin[t=45.03,P<0.001],vimentin[t=67,P<0.001])and MHCC-LM3(NC vs.Lv-FSTL5:E-cadherin[t=50,P<0.001],vimentin[t=72.75,P<0.001])cells at mRNA level.The same as protein level.In addition,the over-expression of FSTL5 inhibited the proliferation(NC vs.Lv-FSTL5:SK-Hep1,3 d[t=7.324,P=0.018],4 d[t=6.23,P=0.021],5 d[t=10.21,P=0.003];MHCC-LM3,3 d[t=4.32,P=0.037],4 d[t=7.49,P=0.012],5 d[t=9.3661,P=0.009])and invasion(NC vs.Lv-FSTL5:SK-Hep1,t=21.57,P<0.001;MHCC-LM3,t=18.04,P<0.001)of HCC cells.Conclusions:Down-regulation of FSTL5 may contribute to EMT of HCC,and FSTL5 is a potential target in the treatment of HCC.展开更多
BACKGROUND Mounting studies have highlighted the pivotal influence of anti-silencing function 1B(ASF1B)on the malignancy of cancers.AIM To explore the influence and mechanism of ASF1B in colorectal cancer(CRC).METHODS...BACKGROUND Mounting studies have highlighted the pivotal influence of anti-silencing function 1B(ASF1B)on the malignancy of cancers.AIM To explore the influence and mechanism of ASF1B in colorectal cancer(CRC).METHODS Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect mRNA expression of ASF1B.Immunohistochemical staining was performed to detect protein expression of ASF1B and Ki67 in tumor tissues.Western blot analysis was used to determine levels of ASF1B and proliferation/epithelial mesenchymal transition(EMT)/stemness-related proteins.In addition,the proliferation of CRC cells was assessed using Cell Counting Kit-8 and 5-Ethynyl-2’-Deoxyuridine assays.The migration and invasion of CRC cells were evaluated using transwell assays.Stemness of CRC cells was tested using the sphere formation assay.To construct a xenograft tumor model,HCT116 cells were introduced into mouse flanks via subcutaneous injection.RESULTS ASF1B expression was markedly increased in CRC tissues and cells,and it was inversely correlated with overall survival of CRC patients and was positively associated with the tumor node metastasis(TNM)stage of CRC patients.Silencing of ASF1B suppressed proliferation,migration,invasion,stemness and EMT of CRC cells as well as tumorigenesis of xenograft mice.Furthermore,protein levels of Pphosphatidylinositol 3-kinase(p-PI3K)and p-AKT were decreased after silencing of ASF1B in CRC cells.The inhibitory effects of ASF1B knockdown on cell proliferation,stemness and EMT were partly abolished by PI3K activator in CRC cells.CONCLUSION Silencing of ASF1B inactivated the PI3K/AKT pathway to suppress CRC malignancy in vitro.展开更多
Colorectal cancer(CRC) is a devastating disease, mainly because of metastasis. As a result, there is a need to better understand the molecular basis of invasion and metastasis and to identify new biomarkers and therap...Colorectal cancer(CRC) is a devastating disease, mainly because of metastasis. As a result, there is a need to better understand the molecular basis of invasion and metastasis and to identify new biomarkers and therapeutic targets to aid in managing these tumors. The actin cytoskeleton and actin-binding proteins are known to play an important role in the process of cancer metastasis because they control and execute essential steps in cell motility and contractility as well as cell division. Caldesmon(CaD) is an actin-binding protein encoded by the CALD1 gene as multiple transcripts that mainly encode two protein isoforms: High-molecular-weight CaD, expressed in smooth muscle, and low-molecular weight CaD(l-CaD), expressed in nonsmooth muscle cells. According to our comprehensive review of the literature, CaD, particularly l-CaD, plays a key role in the development, metastasis, and resistance to chemoradiotherapy in colorectal, breast, and urinary bladder cancers and gliomas, among other malignancies. CaD is involved in many aspects of the carcinogenic hallmarks, including epithelial mesenchymal transition via transforming growth factor-beta signaling, angiogenesis, resistance to hormonal therapy, and immune evasion. Recent data show that CaD is expressed in tumor cells as well as in stromal cells, such as cancerassociated fibroblasts, where it modulates the tumor microenvironment to favor the tumor. Interestingly, CaD undergoes selective tumor-specific splicing, and the resulting isoforms are generally not expressed in normal tissues, making these transcripts ideal targets for drug design. In this review, we will analyze these features of CaD with a focus on CRC and show how the currently available data qualify CaD as a potential candidate for targeted therapy in addition to its role in the diagnosis and prognosis of cancer.展开更多
Objective:Clear cell renal cell carcinoma(ccRCC)is the most common subtype of renal cell carcinoma(RCC)and is characterized by biallelic inactivation of the von Hippel-Lindau(VHL)tumor suppressor gene.One effect of VH...Objective:Clear cell renal cell carcinoma(ccRCC)is the most common subtype of renal cell carcinoma(RCC)and is characterized by biallelic inactivation of the von Hippel-Lindau(VHL)tumor suppressor gene.One effect of VHL inactivation is hypoxia inducible factor alpha(HIFa)-independent constitutive activation of nuclear factor kappa B(NF-κB)and c-jun N-terminal kinase(JNK).Both NF-κB and JNK drive ccRCC growth and epithelial to mesenchymal transition(EMT).The purpose of this study was to determine the biochemical effects of pomegranate juice extracts(PE)on RCC cell lines.Methods:The pre-clinical effects of PE on NF-κB,JNK,and the EMT phenotype were assayed,including its effect on proliferation,anchorage-independent growth,and invasion of pVHLdeficient RCCs.Results:PE inhibits the NF-κB and JNK pathways and consequently inhibits the EMT phenotype of pVHL-deficient ccRCCs.The effects of PE are concentration-dependent and affect not only biochemical markers of EMT(i.e.,cadherin expression)but also functional manifestations of EMT,such as invasion.These effects are manifested within days of exposure to PE when diluted 2000-fold.Highly dilute concentrations of PE(106 dilution),which do not impact these pathways in the short term,were found to have NF-κB and JNK inhibitory effects and ability to reverse the EMT phenotype following prolonged exposure.Conclusion:These findings suggest that PE may mediate inhibition growth of pVHL-deficient ccRCCs and raises the possibility of its use as a dietary adjunct to managing patients with active surveillance for small,localized,incidentally identified renal tumors so as to avoid more invasive procedures such as nephrectomy.展开更多
AIM:To investigate whether upregulation of apoptosisstimulating p53 protein 2(ASPP2)expression could alleviate the development of proliferative vitreoretinopathy(PVR)in a rat model.METHODS:ASPP2-lentivirus or scramble...AIM:To investigate whether upregulation of apoptosisstimulating p53 protein 2(ASPP2)expression could alleviate the development of proliferative vitreoretinopathy(PVR)in a rat model.METHODS:ASPP2-lentivirus or scrambled-lentivirus were transfected into ARPE-19 cells,followed with measurements of cell cytotoxicity by cell counting kit-8 assay.ASPP2 upregulation was confirmed by Western blotting and immunocytochemistry.Then ARPE-19 cells pretreated with ASPP2-lentivirus were intravitreally injected to Brown Norway rats to induce PVR models.PVR development and retinal function were evaluated by retinal photography and electroretinography,respectively.Finally,epithelial-mesenchymal transition as well as autophagy were investigated in rats’retinas via Western blotting.RESULTS:Protein expression of ASPP2 was significantly upregulated by ASPP2-lentivirus transfection in ARPE-19 cells.The development and progression of PVR were impeded significantly in rats with intravitreal injection of ARPE-19 cells pretreated with ASPP2-lentivirus.Accordingly,retinal functions were less affected and PVR grades were much lower in rats with ASPP2-lentivirus compared to scrambledlentivirus treatment.Moreover,epithelial-mesenchymal transition and autophagy markers were decreased in the retinas of rats treated with ASPP2-lentivirus.CONCLUSION:ASPP2-lentivirus transfected to ARPE-19 cells mitigates the progression of PVR in rat models,which might be partly through reduced autophagy and attenuated epithelial-mesenchymal transition.ASPP2 might stand as a new approach for PVR treatment in the future.展开更多
Renal interstitial fibrosis(RIF)is the crucial pathway in chronic kidney disease(CKD)leading to the end-stage renal failure.However,the underlying mechanism of Shen Qi Wan(SQW)on RIF is not fully understood.In the cur...Renal interstitial fibrosis(RIF)is the crucial pathway in chronic kidney disease(CKD)leading to the end-stage renal failure.However,the underlying mechanism of Shen Qi Wan(SQW)on RIF is not fully understood.In the current study,we investigated the role of Aquaporin 1(AQP1)in SQW on tubular epithelial-to-mesenchymal transition(EMT).A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo.Subsequently,the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown.The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine,increased the protein expression of E-cadherin and AQP1 expression,and decreased the expression of vimentin andα-smooth muscle actin(α-SMA).Similarly,treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells.The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1.AQP1 knockdown also increased the mRNA expression of vimentin andα-SMA,and decreased the expression of E-cadherin.The protein expression of vimentin increased,while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells.These results revealed that AQP1 knockdown promoted EMT.Furthermore,AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells.In sum,SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.展开更多
Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and ...Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and absolute quantitation(iTRAQ)-based quantitative proteomic strategy was used to study the secretomes in conditioned media from HCC cancerous tissues,surrounding noncancerous tissues,and distal noncancerous tissues to identify diagnostic and prognostic biomarkers for HCC.In total,22 and 49 dysregulated secretory proteins were identified in the cancerous and surrounding noncancerous tissues,respectively,compared with the distal noncancerous tissues.Among these proteins,carbonic anhydrase II(CA2)was identified to be significantly upregulated in the secretome of cancerous tissues;correspondingly,the serum concentrations of CA2 were remarkably increased in HCC patients compared with that in normal populations.Interestingly,a significant increase of serum CA2 in recurrent HCC patients after radical resection was also confirmed compared with HCC patients without recurrence,and the serum level of CA2 could act as an independent prognostic factor for time to recurrence and overall survival.Regarding the mechanism,the secreted CA2 enhances the migration and invasion of HCC cells by activating the epithelial mesenchymal transition pathway.Taken together,this study identified a novel biomarker for HCC diagnosis and prognosis,and provided a valuable resource of HCC secretome for investigating serological biomarkers.展开更多
Cancer is the second leading cause of death in the US.Current major treatments for cancer management include surgery,cytotoxic chemotherapy,targeted therapy,radiation therapy,endocrine therapy and immunotherapy.Despit...Cancer is the second leading cause of death in the US.Current major treatments for cancer management include surgery,cytotoxic chemotherapy,targeted therapy,radiation therapy,endocrine therapy and immunotherapy.Despite the endeavors and achievements made in treating cancers during the past decades,resistance to classical chemotherapeutic agents and/or novel targeted drugs continues to be a major problem in cancer therapies.Drug resistance,either existing before treatment(intrinsic)or generated after therapy(acquired),is responsible for most relapses of cancer,one of the major causes of death of the disease.Heterogeneity among patients and tumors,and the versatility of cancer to circumvent therapies make drug resistance more challenging to deal with.Better understanding the mechanisms of drug resistance is required to provide guidance to future cancer treatment and achieve better outcomes.In this review,intrinsic and acquired resistance will be discussed.In addition,new discoveries in mechanisms of drug resistance will be reviewed.Particularly,we will highlight roles of ATP in drug resistance by discussing recent findings of exceptionally high levels of intratumoral extracellular ATP as well as intracellular ATP internalized from extracellular environment.The complexity of drug resistance development suggests that combinational and personalized therapies,which should take ATP into consideration,might provide better strategies and improved efficacy for fighting drug resistance in cancer.展开更多
To investigate the effect and underlying mechanism of adenovirus-mediated antisense ERK2(Adanti-ERK2)gene therapy upon chronic allograft nephropathy(CAN)of rats,male Lewis(LEW,RT11)rats received male Fisher(F344,RT11v...To investigate the effect and underlying mechanism of adenovirus-mediated antisense ERK2(Adanti-ERK2)gene therapy upon chronic allograft nephropathy(CAN)of rats,male Lewis(LEW,RT11)rats received male Fisher(F344,RT11v1)renal allografts.The recipients were divided into three groups:(1)empty control group;(2)vector control group;(3)gene therapy group.All recipients were sacrificed for the grafts and serum analysis at the 24th week after transplantation.Morphometric analysis was used to determine thefibrosis of grafts.Immunohistochemistry was used to detect the expression of E-Cadherin,Vimentin,TβR I and the infiltration of CD4+T lymphocyte,CD8+T lymphocyte and ED-1+monocytes.Enzyme linked immunosorbent assay(ELISA)was used to detect TGF-β1 in serum.The grafts in the empty control group and vector control group showed CAN.There was less E-Cadherin in renal tubular epithelial cells in the empty control group but more Vimentin and TβR I.In the gene therapy group,thefibrosis was ameliorated and fewer T lymphocytes and ED-1+monocytes infiltrated in the interstitium.There was no significant difference in the expression of E-Cadherin between the gene therapy group and normal rats.Compared with the empty control group,the expression of TGF-β1 in the gene therapy group was down-regulated.Adanti-ERK2 gene therapy protects the renal allograft and attenuates graftfibrosis,which may be correlated with a decreased renal tubular epithelial mesenchymal transition,a decreased infiltration of CD4+T lymphocyte,CD8+T lymphocytes and ED-1+monocytes in renal interstitium,and the down-regulated TGF-β1 expression.展开更多
Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)h...Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)have been proposed as important mechanisms underlying TNBC metastasis.CDK9 is highly expressed in breast cancer,including TNBC,where it promotes EMT and induces cancer cell stemness.In this study,we have identified a tetrahydroisoquinoline derivative(compound 1)as a potent and selective CDK9-cyclin T1 inhibitor via virtual screening.Interestingly,by targeting the ATP binding site,compound 1 not only inhibited CDK9 activity but also disrupted the CDK9-cyclin T1 proteineprotein interaction(PPI).Mechanistically,compound 1 reversed EMT and reduced the ratio of CSCs by blocking the CDK9-cyclin T1 interaction,leading to reduced TNBC cell proliferation and migration.To date,compound 1 is the first reported tetrahydroisoquinoline-based CDK9-cyclin T1 ATP-competitive inhibitor that also interferes with the interaction between CDK9 and cyclin T1.Compound 1 may serve as a promising scaffold for developing more selective and potent anti-TNBC agents.Our work also provides insight into the role of the CDK9-cyclin T1 PPI on EMT and CSCs and highlights the feasibility and significance of targeting CDK9 for the treatment of TNBC.展开更多
基金Basic Research Foundation for Universities of the CPC Central Committee(No.2042021KF0081)Innovation Group of Natural Science Foundation of Hubei Province(2020CFA027)。
文摘Objective:To investigate the effects of Eriocitrin on the proliferation and migration of Lung adenocarcinoma(LUAD)cells A549 and H1299,and the mechanism of Epithelial-Mesenchymal Transition(EMT).Methods:The effects of different Eriocitrin on the proliferation of LUAD cells A549 and H1299 were examined by CCK8 method.EMT-associated epithelial calmodulin(E-cadherin and N-cadherin),vimentin,ferroptosis-associated protein SLC7A11,GPX4,FTH were detected by Western Blot and expression of mRNA of EMT marker molecules E-cadherin,N-cadherin,Snail were detected by qRT-PCR.Effects of saccharomyces cerevisiae suberin on ferroptosis in LUAD cells as observed by lipid reactive oxygen species(ROS)assay.Results:Eriocitrin could significantly inhibit the proliferative behavior of LUAD cells A549 and H1299 and showed a certain dose-and time-dependence.Compared with the control group,different concentrations of Eriocitrin could significantly reduce the scratch healing rate after 24 and 48 h of action,and the difference was statistically significant(P<0.01).The expression of ROS is increased,EMT-related protein E-cadherin was increased in LUAD cells A549 and H1299 compared with the control group after the intervention with Eriocitrin.N-cadherin and Vimentin expression was decreased.E-cadherin mRNA expression was increased,and N-cadherin,Snail mRNA expression was decreased,expression of ferroptosis-associated protein SLC7A11,GPX4,FTH was decreased,the difference was statistically significant(P<0.05).Conclusion:Eriocitrin may inhibit the proliferation and migration of LUAD cells by regulating the EMT pathway and has potential application in LUAD prevention and adjuvant chemotherapy.
基金Supported by the National Natural Science Foundation of China(No.81072221)National Science Foundation of Hunan Province(No.14JJ2005)
文摘AIM:To analyze the relationship between clinical features and epithelial mesenchymal transition(EMT) in retinoblastoma(RB),further to investigate whether miR-200c regulates the EMT and migration of RB cells.METHODS: Expression of EMT-related markers and tumorrelated factors were detected by immuno-histochemistry analysis in RB tissue from 29 cases. Correlations between their expression and clinical characteristics were analyzed. The regulation effects of miR-200c on EMT-related markers,tumor-related factors were observed in m RNA level and protein level by real-time polymerase chain reaction(PCR) and Western blot,respectively,in Y79 and Weri-rb1 cells. Its effects on migration force of these RB cell lines were also detected with Transwell test.RESULTS: Lower expression of E-cadherin was present in the cases with malignant prognosis. Mi R-200c promoted the expression of E-cadherin and decreased the expression of Vimentin and N-cadherin in Y79 and Weri-rb1 cells. Migration force of RB cells could be inhibited by miR-200c.CONCLUSION: EMT might be associated with bad prognosis in RB. Mi R-200c suppresses the migration of retinoblastomatous cells by reverse EMT.
文摘Triple Negative Breast Cancer (TNBC) is a malignant form of cancer with very high mortality and morbidity. Epithelial to Mesenchymal Transition (EMT) is the most common pathophysiological change observed in cancer cells of epithelial origin that promotes metastasis, drug resistance and cancer stem cell formation. Since the information regarding differential gene expression in TNBC cells and cell signaling events leading to EMT is limited, this investigation was done by comparing transcriptomic data generated by RNA isolation and sequencing of a EMT model TNBC cell line in comparison to regular TNBC cells. RNA sequencing and Ingenuity Pathway Software Analysis (IPA) of the transcriptomic data revealed several upregulated and downregulated gene expressions along with novel core canonical pathways including Sirtuin signaling, Oxidative Phosphorylation and Mitochondrial dysfunction events involved in EMT changes of the TNBC cells.
基金Supported by the State Administration of Traditional Chinese Medicine of China(2009-No.30)
文摘Objective: To examine the effect of brucine on the migration, invasion, adhesion and expressions of epithelial-to-mesenchymal transition(EMT) markers and matrix metalloproteinases(MMPs) in the highly metastatic breast cancer cell lines MDA-MB-231 and Hs578-T. Methods: MDA-MB-231 and Hs578-T cells were divided to 4 groups: the control group(0.1% DMSO), and 25, 50 and 100 μmol/L brucine groups. The cell viability was determined using a Cell Titer-Glo? luminescent cell viability. The scratch wound healing assay and tanswell migration assay were used to determine the migration ability of these cells treated by different concentrations of brucine. The proliferation rate, invasive potential and adhesive ability were respectively performed by colony formation assay, transwell invasion assay and adhension assay. The protein and m RNA expressions of EMT biomarkers, MMP-2 and MMP-9 were investigated by real-time reverse transcription polymerase chain reaction and Western blot. Results: Compared with the control group, brucine had little effect on cell viability or proliferation(P>0.05), but led to a dose-dependent decrease on migration, invasion, adhension of MDA-MB-231 and Hs578-T cells(P<0.01). Furthermore, brucine increased the protein and m RNA levels of EMT markers such as E-cadherin and β-catenin in MDA-MB-231 and Hs578-T cells, and decreased the protein and m RNA levels of mesenychmal markers such as vimentin and fibronectin, as well as the expressions of MMP-2 and MMP-9(all P<0.01). Conclusion: Brucine inhibited triple negative breast cancer cells metastasis potentially through EMT reversion and MMP-2 and MMP-9 inhibition.
文摘BACKGROUND Currently,there are many therapeutic methods for lung adenocarcinoma(LUAD),but the 5-year survival rate is still only 15%at later stages.Epithelial–mesenchymal transition(EMT)has been shown to be closely associated with local dissemination and subsequent metastasis of solid tumors.However,the role of EMT in the occurrence and development of LUAD remains unclear.AIM To further elucidate the value of EMT-related genes in LUAD prognosis.METHODS Univariate,least absolute shrinkage and selection operator,and multivariate Cox regression analyses were applied to establish and validate a new EMT-related gene signature for predicting LUAD prognosis.The risk model was evaluated by Kaplan–Meier survival analysis,principal component analysis,and functional enrichment analysis and was used for nomogram construction.The potential structures of drugs to which LUAD is sensitive were discussed with respect to EMT-related genes in this model.RESULTS Thirty-three differentially expressed genes related to EMT were found to be highly associated with overall survival(OS)by using univariate Cox regression analysis(log2FC≥1,false discovery rate<0.001).A prognostic signature of 7EMT-associated genes was developed to divide patients into two risk groups by high or low risk scores.Kaplan–Meier survival analysis showed that the OS of patients in the high-risk group was significantly poorer than that of patients in the low-risk group(P<0.05).Multivariate Cox regression analysis showed that the risk score was an independent risk factor for OS(HR>1,P<0.05).The results of receiver operator characteristic curve analysis suggested that the 7-gene signature had a perfect ability to predict prognosis(all area under the curves>0.5).CONCLUSION The EMT-associated gene signature classifier could be used as a feasible indicator for predicting OS.
文摘<strong>Objective:</strong> Upfront docetaxel use for hormone na<span style="white-space:nowrap;">ï</span>ve advanced prostate cancer is reported that it successfully delayed the progression to hormone refractory stage, though the adequate methodology to obtain the maximum effect is unclear. We investigated these issues from our experiences of upfront docetaxel use with LH-RH antagonist for metastatic hormone sensitive prostate cancer, aiming at the prevention of epithelial-mesenchymal transition (EMT) for apoptosis tolerance. <strong>Patients and Methods:</strong> Of 31 stage IV new prostate cancer patients treated with upfront docetaxel and LH-RH antagonist (Degarelix), 25 patients who could be followed more than 12 months (mean 36.2 months) were analyzed. Docetaxel was used two to three courses basically 75 mg/m2 dose initializing two weeks after the induction of first Degarelix. <strong>Results:</strong> The clinical course was divided clearly to two groups according to prostate specific antigen (PSA) values. Of 25 patients, 12 patient’s PSA did not decrease below 0.1 ng/ml within 6 months (group A) and gradually rose afterwards. PSA in another 13 patients (group B) decreased below 0.1 within 6 months and kept below 0.1 during the follow up period. Although statistically not significant, the initial group A’s PSA was higher than group B’s (average 1308 and 353 ng/ml), however, number of metastasis, Gleason sum, and bone metastatic extent of disease showed no difference between them. Among group B patients, 7 cases had only upfront docetaxel and hormonal therapy, and some of these patients showed only atrophic gland and fibrotic tissue at second prostate biopsy (specimens after more than two years of therapy), suggesting complete response. <strong>Conclusion:</strong> Our study suggested that PSA value at 6 months may predict the outcome of whole therapy. Patients showing PSA less than 0.1 ng/ml at 6 months and requiring no therapy other than docetaxel and hormone may be induced to complete response. Upfront docetaxel with LH-RH antagonist may prevent EMT for obtaining apoptosis tolerance, in case the patient does not have the castration-resistant clone at the beginning of the therapy (group B).
基金the Natural Science Foundation of Shaanxi Province(No.2022JM-521)the Science and Technology Plan Project of Xi’an(No.21YXYJ0031).
文摘AIM:To explore the effects of hepatocyte growth factor(HGF)on retinal pigment epithelium(RPE)cell behaviors.METHODS:The human adult retinal pigment epithelial cell line-19(ARPE-19)were treated by HGF or mesenchymalepithelial transition factor(MET)inhibitor SU11274 in vitro.Cell viability was detected by a Cell Counting Kit-8 assay.Cell proliferation and motility was detected by a bromodeoxyuridine incorporation assay and a wound healing assay,respectively.The expression levels of MET,phosphorylated MET,protein kinase B(AKT),and phosphorylated AKT proteins were determined by Western blot assay.The MET and phosphorylated MET proteins were also determined by immunofluorescence assay.RESULTS:HGF increased ARPE-19 cells’viability,proliferation and migration,and induced an increase of phosphorylated MET and phosphorylated AKT proteins.SU11274 significantly reduced cell viability,proliferation,and migration and decreased the expression of MET and AKT proteins.SU11274 suppressed HGF-induced increase of viability,proliferation,and migration in ARPE-19 cells.Additionally,SU11274 also blocked HGF-induced phosphorylation of MET and AKT proteins.CONCLUSION:HGF enhances cellular viability,proliferation,and migration in RPE cells through the MET/AKT signaling pathway,whereas this enhancement is suppressed by the MET inhibitor SU11274.HGF-induced MET/AKT signaling might be a vital contributor of RPE cells survival.
基金supported by the Health Fellowship Foundation,Seoul,Korea.
文摘Background:Keloid scarring is a fibroproliferative disease caused by aberrant genetic activation with an unclear underlying mechanism.Genetic predisposition,aberrant cellular responses to environmental factors,increased inflammatory cytokines and epithelial-mesenchymal transition(EMT)phenomena are known as major contributors.In this study,we aimed to identify the molecular drivers that initiate keloid pathogenesis.Methods:Bulk tissue RNA sequencing analyses of keloid and normal tissues along with ex vivo and in vitro tests were performed to identify the contributing genes to keloid pathogenesis.An animal model of inflammatory keloid scarring was reproduced by replication of a skin fibrosis model with intradermal bleomycin injection in C57BL/6 mice.Results:Gene set enrichment analysis revealed upregulation of Wnt family member 5A(WNT5A)expression and genes associated with EMT in keloid tissues.Consistently,human keloid tissues and the bleomycin-induced skin fibrosis animal model showed significantly increased expression ofWNT5A and EMT markers.Increased activation of the interleukin(IL)-6/Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway and subsequent elevation of EMT markerswas also observed in keratinocytes co-cultured withWNT5A-activated fibroblasts or keloid fibroblasts.Furthermore,WNT5A silencing and the blockage of IL-6 secretion via neutralizing IL-6 antibody reversed hyperactivation of the STAT pathway and EMT markers in keratinocytes.Lastly,STAT3 silencing significantly reduced the EMT-like phenotypes in both keratinocytes and IL-6-stimulated keratinocytes.Conclusions:Intercellular communication via the WNT5A and STAT pathways possibly underlies a partial mechanism of EMT-like phenomena in keloid pathogenesis.IL-6 secreted from WNT5A-activated fibroblasts or keloid fibroblasts activates the JAK/STAT signaling pathway in adjacent keratinocytes which in turn express EMT markers.A better understanding of keloid development and the role of WNT5A in EMT will promote the development of next-generation targeted treatments for keloid scars.
基金This study was supported by National Key Clinical Specialties Construction Program of China(No.[2012]649).
文摘SOX4 is highly expressed in gastric cancer(GC)and is associated with tumor grade,metastasis and prognosis,however the mechanism is not clear.We report herein that SOX4 was upregulated and overexpression of SOX4 was associated with increased expression of the markers of Epithelialemesenchymal transition(EMT)and stemness in clinic patient samples.In vitro,overexpression of SOX4 promoted the invasion as showed by Transwell assay and stemness of GC cells as assessed by sphere formation assay,which was suppressed by silencing SOX4 with shRNA.Further studies showed that SOX4 up-regulated the expression of EMT transcription factors Twist1,snail1 and zeb1 and stemness transcription factors SOX2 and OCT4,and promoted the nuclear translocation of β-catenin.Moreover,we revealed that TGF-β treatment significantly up-regulated the expression of SOX4 and silencing SOX4 reversed TGF-β induced invasion and sphere formation ability of GC cells.Finally,we showed that SOX4 promoted the lung metastasis and tumor formation ability of gastric cancer cells in nude mice.Our results suggest that SOX4 is a target TGF-β signaling and mediates TGF-β-induced EMT and stem cell characteristics of GC cells,revealing a novel role of TGF-β/SOX4 axis in the regulation of malignant behavior of GC.
基金National Natural Science Foundation of China(No.81402514)Support Program for Excellent Young Talents in Universities of Anhui Province(No.gxyq2022042)"512 Talent Cultivation Plan" of Bengbu Medical College(No.by51202208)。
文摘Objective: To explore the effect of connexin 32 (Cx32) on the cell proliferation, migration, invasion of human hepatocellular carcinoma (HCC) cell line Huh7 and its mechanism. Methods: Firstly, bioinformatics techniques were used to analyze the difference in expression of Cx32 between HCC tissues and normal liver tissues, and the relationship between Cx32 expression and important clinicopathological features of HCC was also explored. Subsequently, Cx32 expression in HCC cell lines and normal hepatic epithelial cell line was detected in vitro. Huh7 cell line with stable over⁃expression of Cx32 was further established, and the change in cell proliferation ability was measured by MTT assay, changes in migration and invasion capacities were detected by wound⁃healing assay and transwell assay, on this cell line. Finally, western blot and immunofluorescence (IF) were used to investigate the alterations of expression of epithelial⁃mesenchymal transition (EMT) markers. Results: Bioinformatics analyses showed that Cx32 mRNA and protein expression levels in HCC tissues were lower than those in normal liver tissues, and the mRNA expression level of Cx32 was negatively correlated with T stage, histological grade and clinical stage of HCC patients (all P<0.05). Results of in vitro experiments revealed Cx32 protein expression in different HCC cell lines was down⁃regulated compared to that in normal hepatic epithelial cell line LO2. Cx32 stably over⁃expressed (Cx32 OE) Huh7 cell line was successfully constructed by lentivirus infection and showed high expression of Cx32 protein in the cell line. Compared to the control group and (or) the negative control (NC) group, the Cx32 OE group exhibited decreased OD490 value, wound healing rate and invasive cell number (all P<0.05). Furthermore, an increase in the expression of epithelial marker E⁃cadherin, and a decrease in the expression of mesenchymal markers Snail and Vimentin, were observed in Cx32⁃OE Huh7 cell line. Conclusion: Cx32 is low expressed in HCC tissues and cells, while the proliferation, migration and invasion ability of Huh7 cells can be inhibited by over⁃expression of Cx32, of which the underlying mechanism may be related to the inhibition of EMT process.
基金the grants from the Anhui Province Science Foundation for Youths(No.1808085QH288)the University Top-notch Talent Cultivation Project(No.gxbjZD28)+1 种基金the Bengbu Medical College Science Foundation(No.BYKY1734ZD)the Science Foundation of the First Affiliated Hospital of Bengbu Medical College(No.Byyfykj201727).
文摘Background:Epithelial to mesenchymal transition(EMT)is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma(HCC).Follistatin(FST)family members are considered to be an attractive therapeutic targets and prognostic indicators in cancers.As a derivative of FST,Follistatin Like 5(FSTL5)may play a similar role in HCC cells.This study aimed to investigate the expression and function of FSTL5 in HCC and its role in EMT.Methods:FSTL5,E-cadherin and vimentin in HCC,and paracancerous tissues were detected by immunohistochemistry.Correlation of FSTL5 expression with overall survival was assessed.The proliferation and invasion of HCC cell lines SK-Hep1 and MHCC-LM3 were analyzed by cell counting kit-8 and Transwell assays.The expression of FSTL5,E-cadherin,and vimentin in HCC cells was examined by polymerase chain reaction and Western blot analysis.T-test was used to analyze the difference in proliferation and invasion ability between groups.The Spearman rank correlation test was used to detect the correlation between the expression of FSTL5 and E-cadherin or vimentin.Results:The expression of FSTL5 in HCC was lower than that in paracancerous tissues(9.97%vs.82.55%,χ2=340.15,P<0.001).Patients with high FSTL5 expression had a better prognosis(χ2=8.22,P=0.004)and smaller tumor diameter(χ2=45.52,P<0.001),less lymph node metastasis(χ2=5.58,P=0.02),earlier tumor node metastasis stage(χ2=11.29,P=0.001),a reduced number of tumors(χ2=5.05,P=0.02),lower alpha-fetoprotein value(χ2=24.36,P<0.001),more probability of hepatitis carrying(χ2=40.9,P<0.001),and better liver function grade(χ2=5.21,P=0.02).Immunohistochemistry showed that FSTL5 expression in HCC tissues was positively correlated with E-cadherin expression(r=0.38,P<0.001)and negatively correlated with vimentin expression(r=-0.385,P<0.001).Furthermore,over-expression of FSTL5 up-regulated the expression of E-cadherin and down-regulated the expression of vimentin in SK-Hep1(negative control[NC]vs.FSTL5-interfering group[Lv-FSTL5]:E-cadherin[t=45.03,P<0.001],vimentin[t=67,P<0.001])and MHCC-LM3(NC vs.Lv-FSTL5:E-cadherin[t=50,P<0.001],vimentin[t=72.75,P<0.001])cells at mRNA level.The same as protein level.In addition,the over-expression of FSTL5 inhibited the proliferation(NC vs.Lv-FSTL5:SK-Hep1,3 d[t=7.324,P=0.018],4 d[t=6.23,P=0.021],5 d[t=10.21,P=0.003];MHCC-LM3,3 d[t=4.32,P=0.037],4 d[t=7.49,P=0.012],5 d[t=9.3661,P=0.009])and invasion(NC vs.Lv-FSTL5:SK-Hep1,t=21.57,P<0.001;MHCC-LM3,t=18.04,P<0.001)of HCC cells.Conclusions:Down-regulation of FSTL5 may contribute to EMT of HCC,and FSTL5 is a potential target in the treatment of HCC.
基金Huzhou Science and Technology Bureau Foundation,No.2018GY09.
文摘BACKGROUND Mounting studies have highlighted the pivotal influence of anti-silencing function 1B(ASF1B)on the malignancy of cancers.AIM To explore the influence and mechanism of ASF1B in colorectal cancer(CRC).METHODS Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect mRNA expression of ASF1B.Immunohistochemical staining was performed to detect protein expression of ASF1B and Ki67 in tumor tissues.Western blot analysis was used to determine levels of ASF1B and proliferation/epithelial mesenchymal transition(EMT)/stemness-related proteins.In addition,the proliferation of CRC cells was assessed using Cell Counting Kit-8 and 5-Ethynyl-2’-Deoxyuridine assays.The migration and invasion of CRC cells were evaluated using transwell assays.Stemness of CRC cells was tested using the sphere formation assay.To construct a xenograft tumor model,HCT116 cells were introduced into mouse flanks via subcutaneous injection.RESULTS ASF1B expression was markedly increased in CRC tissues and cells,and it was inversely correlated with overall survival of CRC patients and was positively associated with the tumor node metastasis(TNM)stage of CRC patients.Silencing of ASF1B suppressed proliferation,migration,invasion,stemness and EMT of CRC cells as well as tumorigenesis of xenograft mice.Furthermore,protein levels of Pphosphatidylinositol 3-kinase(p-PI3K)and p-AKT were decreased after silencing of ASF1B in CRC cells.The inhibitory effects of ASF1B knockdown on cell proliferation,stemness and EMT were partly abolished by PI3K activator in CRC cells.CONCLUSION Silencing of ASF1B inactivated the PI3K/AKT pathway to suppress CRC malignancy in vitro.
文摘Colorectal cancer(CRC) is a devastating disease, mainly because of metastasis. As a result, there is a need to better understand the molecular basis of invasion and metastasis and to identify new biomarkers and therapeutic targets to aid in managing these tumors. The actin cytoskeleton and actin-binding proteins are known to play an important role in the process of cancer metastasis because they control and execute essential steps in cell motility and contractility as well as cell division. Caldesmon(CaD) is an actin-binding protein encoded by the CALD1 gene as multiple transcripts that mainly encode two protein isoforms: High-molecular-weight CaD, expressed in smooth muscle, and low-molecular weight CaD(l-CaD), expressed in nonsmooth muscle cells. According to our comprehensive review of the literature, CaD, particularly l-CaD, plays a key role in the development, metastasis, and resistance to chemoradiotherapy in colorectal, breast, and urinary bladder cancers and gliomas, among other malignancies. CaD is involved in many aspects of the carcinogenic hallmarks, including epithelial mesenchymal transition via transforming growth factor-beta signaling, angiogenesis, resistance to hormonal therapy, and immune evasion. Recent data show that CaD is expressed in tumor cells as well as in stromal cells, such as cancerassociated fibroblasts, where it modulates the tumor microenvironment to favor the tumor. Interestingly, CaD undergoes selective tumor-specific splicing, and the resulting isoforms are generally not expressed in normal tissues, making these transcripts ideal targets for drug design. In this review, we will analyze these features of CaD with a focus on CRC and show how the currently available data qualify CaD as a potential candidate for targeted therapy in addition to its role in the diagnosis and prognosis of cancer.
文摘Objective:Clear cell renal cell carcinoma(ccRCC)is the most common subtype of renal cell carcinoma(RCC)and is characterized by biallelic inactivation of the von Hippel-Lindau(VHL)tumor suppressor gene.One effect of VHL inactivation is hypoxia inducible factor alpha(HIFa)-independent constitutive activation of nuclear factor kappa B(NF-κB)and c-jun N-terminal kinase(JNK).Both NF-κB and JNK drive ccRCC growth and epithelial to mesenchymal transition(EMT).The purpose of this study was to determine the biochemical effects of pomegranate juice extracts(PE)on RCC cell lines.Methods:The pre-clinical effects of PE on NF-κB,JNK,and the EMT phenotype were assayed,including its effect on proliferation,anchorage-independent growth,and invasion of pVHLdeficient RCCs.Results:PE inhibits the NF-κB and JNK pathways and consequently inhibits the EMT phenotype of pVHL-deficient ccRCCs.The effects of PE are concentration-dependent and affect not only biochemical markers of EMT(i.e.,cadherin expression)but also functional manifestations of EMT,such as invasion.These effects are manifested within days of exposure to PE when diluted 2000-fold.Highly dilute concentrations of PE(106 dilution),which do not impact these pathways in the short term,were found to have NF-κB and JNK inhibitory effects and ability to reverse the EMT phenotype following prolonged exposure.Conclusion:These findings suggest that PE may mediate inhibition growth of pVHL-deficient ccRCCs and raises the possibility of its use as a dietary adjunct to managing patients with active surveillance for small,localized,incidentally identified renal tumors so as to avoid more invasive procedures such as nephrectomy.
基金National Natural Science Foundation of China(No.81800827).
文摘AIM:To investigate whether upregulation of apoptosisstimulating p53 protein 2(ASPP2)expression could alleviate the development of proliferative vitreoretinopathy(PVR)in a rat model.METHODS:ASPP2-lentivirus or scrambled-lentivirus were transfected into ARPE-19 cells,followed with measurements of cell cytotoxicity by cell counting kit-8 assay.ASPP2 upregulation was confirmed by Western blotting and immunocytochemistry.Then ARPE-19 cells pretreated with ASPP2-lentivirus were intravitreally injected to Brown Norway rats to induce PVR models.PVR development and retinal function were evaluated by retinal photography and electroretinography,respectively.Finally,epithelial-mesenchymal transition as well as autophagy were investigated in rats’retinas via Western blotting.RESULTS:Protein expression of ASPP2 was significantly upregulated by ASPP2-lentivirus transfection in ARPE-19 cells.The development and progression of PVR were impeded significantly in rats with intravitreal injection of ARPE-19 cells pretreated with ASPP2-lentivirus.Accordingly,retinal functions were less affected and PVR grades were much lower in rats with ASPP2-lentivirus compared to scrambledlentivirus treatment.Moreover,epithelial-mesenchymal transition and autophagy markers were decreased in the retinas of rats treated with ASPP2-lentivirus.CONCLUSION:ASPP2-lentivirus transfected to ARPE-19 cells mitigates the progression of PVR in rat models,which might be partly through reduced autophagy and attenuated epithelial-mesenchymal transition.ASPP2 might stand as a new approach for PVR treatment in the future.
基金the National Natural Science Foundation of China(Nos.81673839 and 82074304)the Young Talents Foundation of Zhejiang Traditional Chinese Medicine Science and Technology Program(No.2019ZQ014)the Science Foundation of Zhejiang Chinese Medical University(No.2020ZZ13).
文摘Renal interstitial fibrosis(RIF)is the crucial pathway in chronic kidney disease(CKD)leading to the end-stage renal failure.However,the underlying mechanism of Shen Qi Wan(SQW)on RIF is not fully understood.In the current study,we investigated the role of Aquaporin 1(AQP1)in SQW on tubular epithelial-to-mesenchymal transition(EMT).A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo.Subsequently,the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown.The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine,increased the protein expression of E-cadherin and AQP1 expression,and decreased the expression of vimentin andα-smooth muscle actin(α-SMA).Similarly,treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells.The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1.AQP1 knockdown also increased the mRNA expression of vimentin andα-SMA,and decreased the expression of E-cadherin.The protein expression of vimentin increased,while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells.These results revealed that AQP1 knockdown promoted EMT.Furthermore,AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells.In sum,SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.
基金supported by the National Natural Science Foundation of China(Grant Nos.81702910 and 81672376)the Educational Commission of Fujian Province,China(Grant No.2018B013)the Natural Science Foundation of Fujian Province,China(Grant Nos.2019J01298,2017J01159,and 2016J01417)
文摘Early detection and intervention are key strategies to reduce mortality,increase longterm survival,and improve the therapeutic effects of hepatocellular carcinoma(HCC)patients.Herein,the isobaric tag for relative and absolute quantitation(iTRAQ)-based quantitative proteomic strategy was used to study the secretomes in conditioned media from HCC cancerous tissues,surrounding noncancerous tissues,and distal noncancerous tissues to identify diagnostic and prognostic biomarkers for HCC.In total,22 and 49 dysregulated secretory proteins were identified in the cancerous and surrounding noncancerous tissues,respectively,compared with the distal noncancerous tissues.Among these proteins,carbonic anhydrase II(CA2)was identified to be significantly upregulated in the secretome of cancerous tissues;correspondingly,the serum concentrations of CA2 were remarkably increased in HCC patients compared with that in normal populations.Interestingly,a significant increase of serum CA2 in recurrent HCC patients after radical resection was also confirmed compared with HCC patients without recurrence,and the serum level of CA2 could act as an independent prognostic factor for time to recurrence and overall survival.Regarding the mechanism,the secreted CA2 enhances the migration and invasion of HCC cells by activating the epithelial mesenchymal transition pathway.Taken together,this study identified a novel biomarker for HCC diagnosis and prognosis,and provided a valuable resource of HCC secretome for investigating serological biomarkers.
文摘Cancer is the second leading cause of death in the US.Current major treatments for cancer management include surgery,cytotoxic chemotherapy,targeted therapy,radiation therapy,endocrine therapy and immunotherapy.Despite the endeavors and achievements made in treating cancers during the past decades,resistance to classical chemotherapeutic agents and/or novel targeted drugs continues to be a major problem in cancer therapies.Drug resistance,either existing before treatment(intrinsic)or generated after therapy(acquired),is responsible for most relapses of cancer,one of the major causes of death of the disease.Heterogeneity among patients and tumors,and the versatility of cancer to circumvent therapies make drug resistance more challenging to deal with.Better understanding the mechanisms of drug resistance is required to provide guidance to future cancer treatment and achieve better outcomes.In this review,intrinsic and acquired resistance will be discussed.In addition,new discoveries in mechanisms of drug resistance will be reviewed.Particularly,we will highlight roles of ATP in drug resistance by discussing recent findings of exceptionally high levels of intratumoral extracellular ATP as well as intracellular ATP internalized from extracellular environment.The complexity of drug resistance development suggests that combinational and personalized therapies,which should take ATP into consideration,might provide better strategies and improved efficacy for fighting drug resistance in cancer.
基金supported by the National Natural Science Foundation of China(Grant No.30300324).
文摘To investigate the effect and underlying mechanism of adenovirus-mediated antisense ERK2(Adanti-ERK2)gene therapy upon chronic allograft nephropathy(CAN)of rats,male Lewis(LEW,RT11)rats received male Fisher(F344,RT11v1)renal allografts.The recipients were divided into three groups:(1)empty control group;(2)vector control group;(3)gene therapy group.All recipients were sacrificed for the grafts and serum analysis at the 24th week after transplantation.Morphometric analysis was used to determine thefibrosis of grafts.Immunohistochemistry was used to detect the expression of E-Cadherin,Vimentin,TβR I and the infiltration of CD4+T lymphocyte,CD8+T lymphocyte and ED-1+monocytes.Enzyme linked immunosorbent assay(ELISA)was used to detect TGF-β1 in serum.The grafts in the empty control group and vector control group showed CAN.There was less E-Cadherin in renal tubular epithelial cells in the empty control group but more Vimentin and TβR I.In the gene therapy group,thefibrosis was ameliorated and fewer T lymphocytes and ED-1+monocytes infiltrated in the interstitium.There was no significant difference in the expression of E-Cadherin between the gene therapy group and normal rats.Compared with the empty control group,the expression of TGF-β1 in the gene therapy group was down-regulated.Adanti-ERK2 gene therapy protects the renal allograft and attenuates graftfibrosis,which may be correlated with a decreased renal tubular epithelial mesenchymal transition,a decreased infiltration of CD4+T lymphocyte,CD8+T lymphocytes and ED-1+monocytes in renal interstitium,and the down-regulated TGF-β1 expression.
基金This work was supported by the Health and Medical Research Fund(No.HMRF/14150561)the National Natural Science Foundation of China(No.201575121 and 21775131)+3 种基金the Hong Kong Baptist University Century Club Sponsorship Scheme 2020,Teaching Development Fund(No.TDG/1920/02)the Science and Technology Development Fund,Macao SAR,China(File no.0072/2018/A2 and 0007/2020/A1)SKLQRCM(UM)-2020-2022the University of Macao,China(MYRG2019e00002eICMS).
文摘Triple-negative breast cancer(TNBC)is a highly aggressive and metastasizing cancer that has the worst prognosis out of all breast cancer subtypes.The epithelial emesenchymal transition(EMT)and cancer stem cells(CSCs)have been proposed as important mechanisms underlying TNBC metastasis.CDK9 is highly expressed in breast cancer,including TNBC,where it promotes EMT and induces cancer cell stemness.In this study,we have identified a tetrahydroisoquinoline derivative(compound 1)as a potent and selective CDK9-cyclin T1 inhibitor via virtual screening.Interestingly,by targeting the ATP binding site,compound 1 not only inhibited CDK9 activity but also disrupted the CDK9-cyclin T1 proteineprotein interaction(PPI).Mechanistically,compound 1 reversed EMT and reduced the ratio of CSCs by blocking the CDK9-cyclin T1 interaction,leading to reduced TNBC cell proliferation and migration.To date,compound 1 is the first reported tetrahydroisoquinoline-based CDK9-cyclin T1 ATP-competitive inhibitor that also interferes with the interaction between CDK9 and cyclin T1.Compound 1 may serve as a promising scaffold for developing more selective and potent anti-TNBC agents.Our work also provides insight into the role of the CDK9-cyclin T1 PPI on EMT and CSCs and highlights the feasibility and significance of targeting CDK9 for the treatment of TNBC.
