AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was ...AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was determined by real-time reversetranscriptase polymerase chain reaction.Two ErbB2 inhibitory methods,a small molecule ErbB2 kinase inhibitor(AG825) and siRNA,were used to disrupt ErbB2 function in the cell lines.CCA cell invasion,motility and proliferation under ErbB2-disrupted conditions were detected using Transwell and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide assays.In addition,ErbB2 downstream effectors were investigated by Western blotting analysis.RESULTS:Suppression of ErbB2 activity,using a specific kinase inhibitor(AG825),reduced invasion,motility and proliferation of all three CCA cell lines.The ability of this drug to inhibit neoplastic properties(invasion,motility and proliferation) increased concomitantly with the level of ErbB2 expression.Similarly,knockdown of ErbB2 level by siRNA inhibited cell invasion and proliferation of KKU-M213,a high-ErbB2-expressing cell,better than those of the lower-ErbB2-expressing cells,HuCCA-1 and KKU-100.Thus,both inhibitory methods indicated that there is more ErbB2-dependency for malignancy of the high-ErbB2-expressing cell,KKU-M213,than for that of low-ErbB2-expressing ones.In addition,interrupting ErbB2 activity decreased phosphorylation of AKT and p70S6K,but not extracellular signal-regulated kinase 1/2,in the high-ErbB2-expressing CCA cell line.CONCLUSION:Our data indicated that high ErbB2 expression enhances CCA invasion,motility and proliferation via the AKT/p70S6K pathway,which suggests the possibility of targeting these molecules for CCA therapy.展开更多
AIM:To investigate the role of hepatocyte growth factor(HGF) in cholangiocarcinoma(CCA) cell invasiveness and the mechanisms underlying such cellular responses. METHODS:Effects of HGF on cell invasion and motility wer...AIM:To investigate the role of hepatocyte growth factor(HGF) in cholangiocarcinoma(CCA) cell invasiveness and the mechanisms underlying such cellular responses. METHODS:Effects of HGF on cell invasion and motility were investigated in two human CCA cell lines,HuCCA-1 and KKU-M213,using Transwell in vitro assay.Levels of proteins of interest and their phosphorylated forms were determined by Western blotting.Localization of E-cadherin was analyzed by immunofluorescence staining and visualized under confocal microscope. Activities of matrix degrading enzymes were determined by zymography. RESULTS:Both CCA cell lines expressed higher Met levels than the H69 immortalized cholangiocyte cell line.HGF induced invasion and motility of the cell lines and altered E-cadherin from membrane to cytoplasm localization,but did not affect the levels of secreted matrix metalloproteinase(MMP) -2,MMP-9 andurokinase plasminogen activator,key matrix degrading enzymes involved in cell invasion.Concomitantly,HGF stimulated Akt and extracellular signal-regulated kinase(ERK) 1/2 phosphorylation but with slightly different kinetic profiles in the two cell lines.Inhibition of the phosphoinositide 3-kinase(PI3K) /Akt pathway by the PI3K inhibitor,LY294002,markedly suppressed HGFstimulated invasion of both CCA cell lines,and inhibition of the ERK pathway by U0126 suppressed HGF-induced invasion of the KKU-M213 cell line but had a moderate effect on HuCCA-1 cells. CONCLUSION:These data indicate that HGF promotes CCA cell invasiveness through dys-localization of E-cadherin and induction of cell motility by distinct signaling pathways depending on cell line type.展开更多
Objective: Recent studies have shown that tumor-associated macrophages(TAMs) play an important role in cancer invasion and metastasis. Our previous studies have reported that TAMs promote the invasion and metastasis o...Objective: Recent studies have shown that tumor-associated macrophages(TAMs) play an important role in cancer invasion and metastasis. Our previous studies have reported that TAMs promote the invasion and metastasis of gastric cancer(GC) cells through the Kindlin-2 pathway. However, the mechanism needs to be clarified.Methods: THP-1 monocytes were induced by PMA/interleukin(IL)-4/IL-13 to establish an efficient TAM model in vitro and M2 macrophages were isolated via flow cytometry. A dual luciferase reporter system and chromatin immunoprecipitation(Ch IP) assay were used to investigate the mechanism of transforming growth factor β2(TGFβ2) regulating Kindlin-2 expression. Immunohistochemistry was used to study the relationships among TAM infiltration in human GC tissues, Kindlin-2 protein expression, clinicopathological parameters and prognosis in human GC tissues. A nude mouse oncogenesis model was used to verify the invasion and metastasis mechanisms in vivo.Results: We found that Kindlin-2 expression was upregulated at both m RNA and protein levels in GC cells cocultured with TAMs, associated with higher invasion rate. Kindlin-2 knockdown reduced the invasion rate of GC cells under coculture condition. TGFβ2 secreted by TAMs regulated the expression of Kindlin-2 through the transcription factor NF-кB. TAMs thus participated in the progression of GC through the TGFβ2/NF-κB/Kindlin-2 axis. Kindlin-2 expression and TAM infiltration were significantly positively correlated with TNM stage, and patients with high Kindlin-2 expression had significantly poorer overall survival than patients with low Kindlin-2 expression. Furthermore, Kindlin-2 promoted the invasion of GC cells in vivo.Conclusions: This study elucidates the mechanism of TAMs participating in GC cell invasion and metastasis through the TGFβ2/NF-κB/Kindlin-2 axis, providing a possibility for new treatment options and approaches.展开更多
AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of f...AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.展开更多
AIM:To investigate the effects of RNA interference tar-geting hepatocyte progenitor kinase-like kinase(HGK) in the invasion and adhesion of hepatocellular carcinoma(HCC) cell line HepG2.METHODS:Three paired insert DNA...AIM:To investigate the effects of RNA interference tar-geting hepatocyte progenitor kinase-like kinase(HGK) in the invasion and adhesion of hepatocellular carcinoma(HCC) cell line HepG2.METHODS:Three paired insert DNA fragments specif ic to HGK gene and one negative control DNA fragment were synthesized and inserted into RNAi-Ready pSIREN-RetroQ-ZsGreen vector.Western blotting assay and real-time reverse transcriptase polymerase chain reaction(RT-PCR) were used to screen the vector with a highest inhibitory rate.The vector was used to generate recom-binant retrovirus specif ic to HGK.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2h-tetrazolium bromide(MTT) assay was used to examine cell growth;wound closure assay and cell adhesion assay were employed to investigate cell migration and adhesion respectively;and transwell assay and three-dimensional culture invasion assay were used to detect cell invasion.The expressions of matrix metalloproteinase(MMP)-2,MMP-9 and nuclear factor(NF)-κB were detected by Western blotting assay.RESULTS:The real time RT-PCR and Western blotting assay showed that cells transfected with retrovirus me-diating RNAi targeting of HGK(RV-shHGK)-1 vector had the strongest inhibition of HGK protein,with an inhibi-tion rate of 76%,and this vector was used to generate recombinant retrovirus RV-shHGK-1.Cell adhesion assay and MTT assay found that cell adhesion and growth of the cells infected with RV-shHGK-1 were significantly lower than those of the control cells(P < 0.05).Wound closure assay,transwell assay and three-dimensional culture invasion assay showed that the cell invasiveness was significantly less in HGK knockdown cells than in the control cells(P < 0.05).The expressions of MMP-2,MMP-9 and NF-κB were inhibited in HepG2 cells infected with RV-shHGK-1.CONCLUSION:Down-regulation of HGK can obviously inhibit the migration and invasion of HepG2 cells in vitro.HGK may be a new therapeutic target for treatment of HCC.展开更多
Background: The outcomes for patients with advanced hepatocellular carcinoma(HCC) receiving sorafenib are far from satisfactory because of treatment resistance to sorafenib. However, the exact mechanism of resistance ...Background: The outcomes for patients with advanced hepatocellular carcinoma(HCC) receiving sorafenib are far from satisfactory because of treatment resistance to sorafenib. However, the exact mechanism of resistance to sorafenib remains unclear and it is valuable to establish a novel mouse model to quantitatively analyze the inhibition rates of sorafenib on the invasive growth of HCC cells in the liver.Methods: HCC tissue microblocks derived from patients were cultured and mixed with hydrogel drops. Then, hydrogel drops containing microblocks of HCC tissue were attached onto the surface of the livers of nude mice to form lesions or nodules of HCC. The mice received molecular targeting agents through oral administration. Livers with tumor nodules were harvested for H&E staining(hematoxylin-eosin staining) analysis and H&E staining images were quantitatively analyzed using image J software. The invasive growth of HCC cells into the liver was calculated using the depth of the lesions compared with the total thickness of the liver.Results: Microblocks containing cells derived from HCC patients can form lesions in the liver of nude mice. Oral administration of molecular targeting agents inhibited the invasive growth of HCC cells in the liver of nude mice.Conclusions: The model established in this study involves the invasive growth of HCC cells in the liver of nude mice, and the model allows for the quantitative analysis of the inhibitory effect of molecular targeting agents on the invasion of HCC cells in vivo.展开更多
Exercise benefits the musculoskeletal system and reduces the effects of cancer.The effects of exercise are multifactorial,where metabolic changes and tissue adaptation influence outcomes.Mechanical signals,a principal...Exercise benefits the musculoskeletal system and reduces the effects of cancer.The effects of exercise are multifactorial,where metabolic changes and tissue adaptation influence outcomes.Mechanical signals,a principal component of exercise,are anabolic to the musculoskeletal system and restrict cancer progression.We examined the mechanisms through which cancer cells sense and respond to low-magnitude mechanical signals introduced in the form of vibration.Low-magnitude,high-frequency vibration was applied to human breast cancer cells in the form of low-intensity vibration(LIV).LIV decreased matrix invasion and impaired secretion of osteolytic factors PTHLH,IL-11,and RANKL.Furthermore,paracrine signals from mechanically stimulated cancer cells,reduced osteoclast differentiation and resorptive capacity.Disconnecting the nucleus by knockdown of SUN1 and SUN2 impaired LIV-mediated suppression of invasion and osteolytic factor secretion.LIV increased cell stiffness;an effect dependent on the LINC complex.These data show that mechanical vibration reduces the metastatic potential of human breast cancer cells,where the nucleus serves as a mechanosensory apparatus to alter cell structure and intercellular signaling.展开更多
AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression a...AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.METHODS:Liver samples from 36 patients(who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1(TGF-β1) or CCN2 mRNA by in situ hybridization.Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma.Fibroblast-specific protein-1(FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition,α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells,and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining.CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.RESULTS:In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci.In comparison to normal controls,CCN2 mRNA was enhanced 1.9-fold in carcinoma foci(12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma(60.27 ± 28.71 vs 6.42 ± 2.35),with concomitant expression of CCN2 and TGF-β1 mRNA in those areas.Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36(33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature.Incubation of HepG2 cells with CCN2(100 ng/mL) resulted in more of the cells transitioning into S phase(23.85 ± 2.35 vs 10.94 ± 0.23),and induced a significant migratory(4.0-fold) and invasive(5.7-fold) effect.TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.CONCLUSION:These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.展开更多
Objective: To study the effect of laparoscopic surgery and laparotomy on oxidative stress response and cell invasion in lesions after hysteromyomectomy. Methods: Patients with uterine fibroids who received surgical re...Objective: To study the effect of laparoscopic surgery and laparotomy on oxidative stress response and cell invasion in lesions after hysteromyomectomy. Methods: Patients with uterine fibroids who received surgical resection in our hospital between August 2014 and March 2017 were retrospectively analyzed and divided into laparoscopy group and laparotomy group according to different surgical procedures. Immediately after surgery and 24 h after surgery, the contents of oxidative stress response indexes in serum were measured;after surgical resection, the uterine fibroid lesion was collected to determine the expression of cell invasion indexes. Results: Serum MDA, NE, E, Cor and ACTH levels of laparoscopy group immediately after surgery and 24 h after surgery were significantly lower than those of laparotomy group while GSH-Px and T-SOD levels were significantly higher than those of laparotomy group;after surgical resection, CXCL12, CXCR4, MMP2, MMP7 and MMP9 mRNA expression in uterine fibroid lesions of laparoscopy group were significantly lower than those of laparotomy group while TIMP1, TIMP2, RECK and E-cadherin mRNA expression were significantly higher than those of laparotomy group. Conclusion: Laparoscopic surgery can reduce the oxidative stress response after hysteromyoma and inhibit the invasive growth of cells in the lesion.展开更多
AIM:To profile expression of micro RNAs(mi RNAs)in gastric cancer cells and investigate the effect of mi R-374b-5p on gastric cancer cell invasion and metastasis.METHODS:An mi RNA microarray assay was performed to ide...AIM:To profile expression of micro RNAs(mi RNAs)in gastric cancer cells and investigate the effect of mi R-374b-5p on gastric cancer cell invasion and metastasis.METHODS:An mi RNA microarray assay was performed to identify mi RNAs differentially expressed in gastric cancer cell lines(MGC-803 and SGC-7901)compared with a normal gastric epithelial cell line.Upregulation of mi R-374b-5p was newly identified and confirmed via quantitative real-time reverse transcriptionPCR(q RT-PCR).MGC-803 cells were transfected with a synthesized anti-mi R-374b-5p sequence or a control vector using Lipofectamine reagent,or treated with transfection reagent alone or phosphate-buffered saline as controls.Rate of transfection was verified after 48 h by q RT-PCR.Cells were then subjected to transwell migration,wound scratch and cell counting kit-8 assays.A bioinformatic analysis to identify mi R-374b-5p target genes was performed using mi Randa,Pic Tar and Target Scan software.A dual luciferase reporter assay was performed to evaluate the influence of mi R-374b-5p on target gene activation,and q RT-PCR and Western blot were used to evaluate the levels of target m RNA and protein following transfection with mi R-374b-5p antisense oligonucleotides.RESULTS:The microarray profiling revealed downregulation of 14(fold change<0.667;P<0.05)and upregulation of 12(fold change>1.50;P<0.05)mi RNAs in MGC-803 and SGC-7901 cells compared with GES-1 controls.The upregulation of mi R-374b-5p(fold change=1.75 and 1.64 in MGC-803 and SGC-7901,respectively;P<0.05)was confirmed by q RT-PCR.Compared with the control groups,the restoration of mi R-374b-5p expression with anti-mi R-374b-5p significantly suppressed the metastasis,invasion and proliferation of MGC-803 cells.The bioinformatic analysis predicted that the 3’untranslated region(UTR)of reversion-inducing cysteine-rich protein with Kazal motif(RECK)contains three mi R-374b-5p target sequences.RECK was verified as a target gene in a dual luciferase reporter assay showing that activation of RECK 3’UTR-pmir GLO was increased by co-transfection with mi R-374b-5p.Finally,transfection of mi R-374b-5p antisense oligonucleotides increased m RNA and protein levels of RECK in MGC-803cells(P<0.05).CONCLUSION:These findings indicate that upregulation of mi R-374b-5p contributes to gastric cancer cell metastasis and invasion through inhibition of RECK expression.展开更多
AIM: To investigate RNA interference targeting signal transducer and activator of transcription-3 (STAT3) on invasion of human pancreatic cancer cells. METHODS: We constructed three plasmids of RNA interference target...AIM: To investigate RNA interference targeting signal transducer and activator of transcription-3 (STAT3) on invasion of human pancreatic cancer cells. METHODS: We constructed three plasmids of RNA interference targeting the STAT3 gene. After LV (lentivirus)-STAT3siRNA (STAT3 small interfering RNA) the vector was transfected into the human pancreatic cell line, SW1990 and cell proliferation was measured by the MTT assay. Flow cytometry was used to assess cell cycle. Vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) mRNA and protein expression were examined by quantitative PCR and western blotting, respectively. The invasion ability of SW1990 cells was determined by cell invasion assay. RESULTS: We successfully constructed the LVSTAT3siRNA lentivirus vector and proved that it can suppress expression of STAT3 gene in SW1990 cells. RNA interference of STAT3 by the LV-STAT3siRNA construct signifi cantly inhibited the growth of SW1990 cells, in addition to signifi cantly decreasing both VEGF and MMP-2 mRNA and protein expression. Moreover, suppression of STAT3 by LV-STAT3siRNA decreased the invasion ability of SW1990 cells.CONCLUSION: The STAT3 signaling pathway may provide a novel therapeutic target for the treatment of pancreatic cancer since it inhibits the invasion ability of pancreatic cancer cells.展开更多
Objective:To study the correlation of c-fos and p16 expression in oral squamous cell carcinoma with cell cycle and cell invasion.Methods: Patients with oral squamous cell carcinoma who underwent surgical resection in ...Objective:To study the correlation of c-fos and p16 expression in oral squamous cell carcinoma with cell cycle and cell invasion.Methods: Patients with oral squamous cell carcinoma who underwent surgical resection in Zaozhuang Mining Group Central Hospital between June 2014 and March 2017 were selected as OSCC group of the research, and patients who received impacted tooth extraction and provided normal gingival mucosa tissue in the Zaozhuang Mining Group Central Hospital during the same period were selected as the control group. The lesion tissue and normal tissue were obtained from OSCC group and control group respectively to determine the expression of c-fos, p16, cell cycle molecules and cell invasion molecules.Results: c-fos mRNA expression in lesion tissue of OSCC group was significantly higher than that of control group, p16 mRNA expression was significantly lower than that of control group, and the c-fos mRNA expression was negatively correlated with p16 mRNA expression;Chk1 and Chk2 mRNA expression in lesion tissue of OSCC group were significantly lower than those of control group whereas NEK2, CyclinD1, MALAT1, Periostin,β-catenin and Vimentin mRNA expression were significantly higher than those of control group;Chk1 and Chk2 mRNA expression in OSCC lesion tissue with high c-fos expression were significantly lower than those in OSCC lesion tissue with low c-fos expression whereas NEK2, CyclinD1, MALAT1, Periostin,β-catenin and Vimentin mRNA expression were significantly higher than those in OSCC lesion tissue with low c-fos expression.Conclusion:The high expression of c-fos and the low expression of p16 in oral squamous cell carcinoma can accelerate cell cycle and promote cell invasion.展开更多
Objective:To study the correlation of miR-106b in nasopharyngeal carcinoma with cell cycle progression and cell invasion.Methods: Patients who were diagnosed with nasopharyngeal carcinoma by pathological biopsy in She...Objective:To study the correlation of miR-106b in nasopharyngeal carcinoma with cell cycle progression and cell invasion.Methods: Patients who were diagnosed with nasopharyngeal carcinoma by pathological biopsy in Shenmu Hospital between March 2013 and February 2018 were included in the study, and the nasopharyngeal carcinoma tissue and tissue adjacent to nasopharyngeal carcinoma obtained from biopsy were collected, miRNA was extracted to determine miR-106b expression, and RNA was extracted to determine the mRNA expression of cell cycle-related genes and cell invasion-related genes.Results: miR-106b expression as well as FOXC1, CyclinD1, CyclinE, TRIM27, uPAR, PARP1 and MTA1 mRNA expression in nasopharyngeal carcinoma tissues were higher than those in adjacent tissues, CYB5R2, p16, IRF5 and E-cadherin mRNA expression were significantly lower than those in adjacent tissues whereas;CYB5R2, p16, IRF5 and E-cadherin mRNA expression in nasopharyngeal carcinoma tissues with high miR-106b expression were lower than those in nasopharyngeal carcinoma tissues with low miR-106b expression whereas FOXC1, CyclinD1, CyclinE, TRIM27, uPAR, PARP1 and MTA1 mRNA expression were higher than those in nasopharyngeal carcinoma tissues with low miR-106b expression.Conclusion:The lowly expressed miR-106b in nasopharyngeal carcinoma can promote the cell cycle progression and cell invasion in the lesion.展开更多
Objective: To study the correlation of S100A13 and FOXA1 expression with cell cycle and cell invasion in fine needle aspiration thyroid carcinoma tissue. Methods: Patients who received ultrasound-guided thyroid nodule...Objective: To study the correlation of S100A13 and FOXA1 expression with cell cycle and cell invasion in fine needle aspiration thyroid carcinoma tissue. Methods: Patients who received ultrasound-guided thyroid nodule fine needle aspiration in Haiyang People's Hospital between April 2015 and February 2017 were selected, and the tissues were divided into malignant thyroid tissue and benign thyroid nodules according to the pathological results after biopsy. The expression of S100A13, FOXA1, cell cycle molecules and cell invasion molecules were measured. Results: S100A13, FOXA1, CDK2, CyclinD1, MCM2, MCM7, SKP2, CLOCK, STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue were significantly higher than those in benign thyroid nodule;CDK2, CyclinD1, MCM2, MCM7, SKP2 and CLOCK mRNA expression in thyroid carcinoma tissue with high FOXA1 expression were significantly higher than those in thyroid carcinoma tissue with low FOXA1 expression;STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue with high S100A13 expression were significantly higher than those in thyroid cancer tissue with low S100A13 expression. Conclusions: High expression of S100A13 and FOXA1 in thyroid carcinoma can promote cell invasion and cell cycle progression.展开更多
Objective:To study the effects of high-intensity focused ultrasound (HIFU) combined with interventional chemoembolization on the advanced cervical cancer lesion growth and cell invasion.Methods:Patients with stage IIB...Objective:To study the effects of high-intensity focused ultrasound (HIFU) combined with interventional chemoembolization on the advanced cervical cancer lesion growth and cell invasion.Methods:Patients with stage IIB-IVA cervical cancer treated in Suining Hospital of TCM between May 2014 and October 2016 were selected and randomly divided into two groups, HIFU group received HIFU combined with interventional chemoembolization, and the control group accepted interventional chemoembolization. The levels of tumor markers in serum as well as the expression of tumor suppressor genes and invasion genes in tumor lesions were determined before and after treatment.Results: 2 weeks, 3 weeks and 4 weeks after treatment, serum TK-1, SCC and CA125 levels of both groups were lower than those before treatment, serum TK-1, SCC and CA125 levels of HIFU group 2 weeks after treatment were not different from those of control group, and serum TK-1, SCC and CA125 levels of HIFU group 3 weeks and 4 weeks after treatment were significantly lower than those of control group;4 weeks after treatment, AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of both groups were higher than those before treatment while FAK, KGFR and MMP9 mRNA expression were lower than those before treatment, and AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of HIFU group were higher than those of control group while FAK, KGFR and MMP9 mRNA expression were lower than those of control group.Conclusion: HIFU combined with interventional chemoembolization can be more effective in suppressing the advanced cervical cancer lesion growth and cell invasion than interventional chemoembolization alone.展开更多
Introduction Hematogenous metastasis is the mainly leading cause of death in breast carcinoma patients.A better understanding of the underlying molecular and cellular mechanisms is crucial for the development of effec...Introduction Hematogenous metastasis is the mainly leading cause of death in breast carcinoma patients.A better understanding of the underlying molecular and cellular mechanisms is crucial for the development of effective treatment for metastatic breast cancer<sup>[</sup>1].It has been well established that cell adhesion and invasion is mediated by a variety of transmembrane proteins,including integrins,cadherins,selectins,and intercellular adhesion molecules.Among these adhesion molecules,the integrins and their downstream signaling pathways have been extensively studied<sup>[2]</sup>.On the other hand,the specific events determining tumor cell interactions with endo-展开更多
Ovarian cancer(OC)is a major cause of cancer-related deaths among gynaecologicalmalignancies.Emerging studies suggest that the long non-coding RNA(lncRNA)may be the potential biomarker for the diagnosis and prognosis ...Ovarian cancer(OC)is a major cause of cancer-related deaths among gynaecologicalmalignancies.Emerging studies suggest that the long non-coding RNA(lncRNA)may be the potential biomarker for the diagnosis and prognosis of the cancer.The current study was carried out to investigate the role of lncRNA CCHE1 silencing in OC cell invasion and migration.Expression of lncRNA CCHE1 in normal ovarian cell Hose and OC cell lines HO 8910,A2780 and SKOV3 was detected.LncRNA were transfected with siRNA,and then the proliferation of cells was detected by using MTT assay.Cell invasion and migration was measured by using Transwell assay and scratch test,respectively.The protein levels of E-cadherin,N-cadherin,ERK,p38-MAPK and the phosphorylation of ERK and p38-MAPK in cells after siRNA transfection were detected by using Western blot analysis.Consequently,lncRNA CCHE1 expression was highly expressed in OC cell lines,especially in SKOV3 cells.siRNA1,siRNA2 and siRNA3 all decreased.lncRNA CCHE1 expression in SKOV3 cells and siRNA2 showed the best silencing efficacy.Silencing of lncRNA CCHE1 decreased proliferation,invasion and migration,and reduced the protein levels of N-cadherin,ERK,p38-MAPK and the phosphorylation of ERK and p38-MAPK,while reducing the protein level of E-cadherin in SKOV3 cells.Collectively,our study proved that the silencing of lncRNA CCHE1 could inhibit SKOV3 cell invasion and migration via inactivating the p38-MAPK signaling pathway.展开更多
Objective:To study the relationship between osteopontin expression and cell invasion as well as angiogenesis in preeclampsia placenta tissue.Methods:A total of 56 patients diagnosed with preeclampsia in our hospital b...Objective:To study the relationship between osteopontin expression and cell invasion as well as angiogenesis in preeclampsia placenta tissue.Methods:A total of 56 patients diagnosed with preeclampsia in our hospital between June 2014 and March 2016 were selected as preeclampsia group (PE group) and 80 normal subjects receiving antenatal examination and giving birth during the same period were selected as control group. After labor, the placenta tissue was collected, immunohistochemical staining kits were used to detect OPN expression and enzyme-linked immunosorbent assay kits were used to determine cell invasion and angiogenesis molecule expression.Results:Positive OPN rate in placenta tissue of PE group was significantly lower than that of control group;cell invasion molecules LAMA4, uPA, Rho and ROCK as well as angiogenesis molecules VEGF, PIGF and AGF protein expression in placenta tissue of PE group were significantly lower than those of control group, and cell invasion molecules LAMA4, uPA, Rho and ROCK as well as angiogenesis molecules VEGF, PIGF and AGF protein expression in OPN-positive placenta tissue were significantly higher than those in OPN-negative placenta tissue.Conclusion:Lowly expressed OPN in preeclampsia placenta tissue will inhibit the expression of cell invasion molecules and angiogenesis molecules so as to inhibit sertoli cell invasion and the angiogenesis in placenta tissue.展开更多
Objective: To study the effect of mesoderm-specific transcript (MEST) on the angiogenesis and trophoblast cell invasion in the preeclampsia placenta. Methods: The puerperae who were diagnosed with preeclampsia and gav...Objective: To study the effect of mesoderm-specific transcript (MEST) on the angiogenesis and trophoblast cell invasion in the preeclampsia placenta. Methods: The puerperae who were diagnosed with preeclampsia and gave birth in our hospital between June 2014 and February 2017 were selected as the PE group, and the healthy puerperae who gave birth in our hospital during the same period were selected as the control group. After delivery, the placental tissue was collected, fluorescence quantitative PCR method was used to determine MEST mRNA expression, and enzyme-linked immunosorbent assay was used to determine the protein levels of angiogenesis molecules and cell invasion molecules. Results: MEST, Notch1, DLL4, VEGF, Rho, ROCK, LAMA4, Notch2 and Notch3 protein levels in placental tissue of PE group were obviously lower than those of control group whereas DNC, sFlt-1 and sEng protein levels were obviously higher than those of control group;Notch1, DLL4, VEGF, Rho, ROCK, LAMA4, Notch2 and Notch3 protein levels in PE placenta tissue with lower MEST expression were greatly lower than those in PE placenta tissue with higher MEST expression whereas DNC, sFlt-1 and sEng protein levels were greatly higher than those in PE placenta tissue with higher MEST expression. Conclusion: The low expression of MEST in preeclampsia placenta can inhibit the angiogenesis and trophoblast cell invasion.展开更多
基金Supported by A grant from Mahidol University,Thailand (to Suthiphongchai T)a scholarship from the Royal Golden Jubilee PhD Program,the Thailand Research Fund (to Treekit-karnmongkol W)
文摘AIM:To compare the impact of ErbB2 on cell invasion and proliferation in cholangiocarcinoma(CCA) cell lines.METHODS:Level of endogenous ErbB2 expression in three CCA cell lines,namely HuCCA-1,KKU-100 and KKU-M213,was determined by real-time reversetranscriptase polymerase chain reaction.Two ErbB2 inhibitory methods,a small molecule ErbB2 kinase inhibitor(AG825) and siRNA,were used to disrupt ErbB2 function in the cell lines.CCA cell invasion,motility and proliferation under ErbB2-disrupted conditions were detected using Transwell and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide assays.In addition,ErbB2 downstream effectors were investigated by Western blotting analysis.RESULTS:Suppression of ErbB2 activity,using a specific kinase inhibitor(AG825),reduced invasion,motility and proliferation of all three CCA cell lines.The ability of this drug to inhibit neoplastic properties(invasion,motility and proliferation) increased concomitantly with the level of ErbB2 expression.Similarly,knockdown of ErbB2 level by siRNA inhibited cell invasion and proliferation of KKU-M213,a high-ErbB2-expressing cell,better than those of the lower-ErbB2-expressing cells,HuCCA-1 and KKU-100.Thus,both inhibitory methods indicated that there is more ErbB2-dependency for malignancy of the high-ErbB2-expressing cell,KKU-M213,than for that of low-ErbB2-expressing ones.In addition,interrupting ErbB2 activity decreased phosphorylation of AKT and p70S6K,but not extracellular signal-regulated kinase 1/2,in the high-ErbB2-expressing CCA cell line.CONCLUSION:Our data indicated that high ErbB2 expression enhances CCA invasion,motility and proliferation via the AKT/p70S6K pathway,which suggests the possibility of targeting these molecules for CCA therapy.
基金Supported by Mahidol University,Thailand and Thailand Research Fund(Suthiphongchai T)Strategic Consortia for Capacity Building of University Faculties and Staff Scholarship,Commission on Higher Education,Ministry of Education,Thailand(Menakongka A)
文摘AIM:To investigate the role of hepatocyte growth factor(HGF) in cholangiocarcinoma(CCA) cell invasiveness and the mechanisms underlying such cellular responses. METHODS:Effects of HGF on cell invasion and motility were investigated in two human CCA cell lines,HuCCA-1 and KKU-M213,using Transwell in vitro assay.Levels of proteins of interest and their phosphorylated forms were determined by Western blotting.Localization of E-cadherin was analyzed by immunofluorescence staining and visualized under confocal microscope. Activities of matrix degrading enzymes were determined by zymography. RESULTS:Both CCA cell lines expressed higher Met levels than the H69 immortalized cholangiocyte cell line.HGF induced invasion and motility of the cell lines and altered E-cadherin from membrane to cytoplasm localization,but did not affect the levels of secreted matrix metalloproteinase(MMP) -2,MMP-9 andurokinase plasminogen activator,key matrix degrading enzymes involved in cell invasion.Concomitantly,HGF stimulated Akt and extracellular signal-regulated kinase(ERK) 1/2 phosphorylation but with slightly different kinetic profiles in the two cell lines.Inhibition of the phosphoinositide 3-kinase(PI3K) /Akt pathway by the PI3K inhibitor,LY294002,markedly suppressed HGFstimulated invasion of both CCA cell lines,and inhibition of the ERK pathway by U0126 suppressed HGF-induced invasion of the KKU-M213 cell line but had a moderate effect on HuCCA-1 cells. CONCLUSION:These data indicate that HGF promotes CCA cell invasiveness through dys-localization of E-cadherin and induction of cell motility by distinct signaling pathways depending on cell line type.
基金supported by grants from the National Natural Science Foundation of China (No. 81372291).
文摘Objective: Recent studies have shown that tumor-associated macrophages(TAMs) play an important role in cancer invasion and metastasis. Our previous studies have reported that TAMs promote the invasion and metastasis of gastric cancer(GC) cells through the Kindlin-2 pathway. However, the mechanism needs to be clarified.Methods: THP-1 monocytes were induced by PMA/interleukin(IL)-4/IL-13 to establish an efficient TAM model in vitro and M2 macrophages were isolated via flow cytometry. A dual luciferase reporter system and chromatin immunoprecipitation(Ch IP) assay were used to investigate the mechanism of transforming growth factor β2(TGFβ2) regulating Kindlin-2 expression. Immunohistochemistry was used to study the relationships among TAM infiltration in human GC tissues, Kindlin-2 protein expression, clinicopathological parameters and prognosis in human GC tissues. A nude mouse oncogenesis model was used to verify the invasion and metastasis mechanisms in vivo.Results: We found that Kindlin-2 expression was upregulated at both m RNA and protein levels in GC cells cocultured with TAMs, associated with higher invasion rate. Kindlin-2 knockdown reduced the invasion rate of GC cells under coculture condition. TGFβ2 secreted by TAMs regulated the expression of Kindlin-2 through the transcription factor NF-кB. TAMs thus participated in the progression of GC through the TGFβ2/NF-κB/Kindlin-2 axis. Kindlin-2 expression and TAM infiltration were significantly positively correlated with TNM stage, and patients with high Kindlin-2 expression had significantly poorer overall survival than patients with low Kindlin-2 expression. Furthermore, Kindlin-2 promoted the invasion of GC cells in vivo.Conclusions: This study elucidates the mechanism of TAMs participating in GC cell invasion and metastasis through the TGFβ2/NF-κB/Kindlin-2 axis, providing a possibility for new treatment options and approaches.
基金Supported by the National Natural Science Foundation of China,No.81201972the China Postdoctoral Science Foundation,No.2013M531555the Postdoctoral Science Foundation of Jiangxi province,No.2013KY44
文摘AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.
基金Supported by The Natural Science Foundation of China,No. 81071692
文摘AIM:To investigate the effects of RNA interference tar-geting hepatocyte progenitor kinase-like kinase(HGK) in the invasion and adhesion of hepatocellular carcinoma(HCC) cell line HepG2.METHODS:Three paired insert DNA fragments specif ic to HGK gene and one negative control DNA fragment were synthesized and inserted into RNAi-Ready pSIREN-RetroQ-ZsGreen vector.Western blotting assay and real-time reverse transcriptase polymerase chain reaction(RT-PCR) were used to screen the vector with a highest inhibitory rate.The vector was used to generate recom-binant retrovirus specif ic to HGK.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2h-tetrazolium bromide(MTT) assay was used to examine cell growth;wound closure assay and cell adhesion assay were employed to investigate cell migration and adhesion respectively;and transwell assay and three-dimensional culture invasion assay were used to detect cell invasion.The expressions of matrix metalloproteinase(MMP)-2,MMP-9 and nuclear factor(NF)-κB were detected by Western blotting assay.RESULTS:The real time RT-PCR and Western blotting assay showed that cells transfected with retrovirus me-diating RNAi targeting of HGK(RV-shHGK)-1 vector had the strongest inhibition of HGK protein,with an inhibi-tion rate of 76%,and this vector was used to generate recombinant retrovirus RV-shHGK-1.Cell adhesion assay and MTT assay found that cell adhesion and growth of the cells infected with RV-shHGK-1 were significantly lower than those of the control cells(P < 0.05).Wound closure assay,transwell assay and three-dimensional culture invasion assay showed that the cell invasiveness was significantly less in HGK knockdown cells than in the control cells(P < 0.05).The expressions of MMP-2,MMP-9 and NF-κB were inhibited in HepG2 cells infected with RV-shHGK-1.CONCLUSION:Down-regulation of HGK can obviously inhibit the migration and invasion of HepG2 cells in vitro.HGK may be a new therapeutic target for treatment of HCC.
基金National Natural Science Foundation of China, Grant/Award Number: 81702986Beijing Municipal Natural Science Foundation, Grant/Award Number: 7142131
文摘Background: The outcomes for patients with advanced hepatocellular carcinoma(HCC) receiving sorafenib are far from satisfactory because of treatment resistance to sorafenib. However, the exact mechanism of resistance to sorafenib remains unclear and it is valuable to establish a novel mouse model to quantitatively analyze the inhibition rates of sorafenib on the invasive growth of HCC cells in the liver.Methods: HCC tissue microblocks derived from patients were cultured and mixed with hydrogel drops. Then, hydrogel drops containing microblocks of HCC tissue were attached onto the surface of the livers of nude mice to form lesions or nodules of HCC. The mice received molecular targeting agents through oral administration. Livers with tumor nodules were harvested for H&E staining(hematoxylin-eosin staining) analysis and H&E staining images were quantitatively analyzed using image J software. The invasive growth of HCC cells into the liver was calculated using the depth of the lesions compared with the total thickness of the liver.Results: Microblocks containing cells derived from HCC patients can form lesions in the liver of nude mice. Oral administration of molecular targeting agents inhibited the invasive growth of HCC cells in the liver of nude mice.Conclusions: The model established in this study involves the invasive growth of HCC cells in the liver of nude mice, and the model allows for the quantitative analysis of the inhibitory effect of molecular targeting agents on the invasion of HCC cells in vivo.
基金Department of Defense BC150678P1(WRT),NIH AR069943-01(WRT),NIH AR068332(US),and Department of Defense BC150678(TAG).
文摘Exercise benefits the musculoskeletal system and reduces the effects of cancer.The effects of exercise are multifactorial,where metabolic changes and tissue adaptation influence outcomes.Mechanical signals,a principal component of exercise,are anabolic to the musculoskeletal system and restrict cancer progression.We examined the mechanisms through which cancer cells sense and respond to low-magnitude mechanical signals introduced in the form of vibration.Low-magnitude,high-frequency vibration was applied to human breast cancer cells in the form of low-intensity vibration(LIV).LIV decreased matrix invasion and impaired secretion of osteolytic factors PTHLH,IL-11,and RANKL.Furthermore,paracrine signals from mechanically stimulated cancer cells,reduced osteoclast differentiation and resorptive capacity.Disconnecting the nucleus by knockdown of SUN1 and SUN2 impaired LIV-mediated suppression of invasion and osteolytic factor secretion.LIV increased cell stiffness;an effect dependent on the LINC complex.These data show that mechanical vibration reduces the metastatic potential of human breast cancer cells,where the nucleus serves as a mechanosensory apparatus to alter cell structure and intercellular signaling.
基金Supported by National Natural Scientific Foundation,No. 30872236,81070370,to Gao RPNIH 5R01AA016003,to Brigstock DR
文摘AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.METHODS:Liver samples from 36 patients(who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1(TGF-β1) or CCN2 mRNA by in situ hybridization.Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma.Fibroblast-specific protein-1(FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition,α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells,and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining.CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.RESULTS:In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci.In comparison to normal controls,CCN2 mRNA was enhanced 1.9-fold in carcinoma foci(12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma(60.27 ± 28.71 vs 6.42 ± 2.35),with concomitant expression of CCN2 and TGF-β1 mRNA in those areas.Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36(33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature.Incubation of HepG2 cells with CCN2(100 ng/mL) resulted in more of the cells transitioning into S phase(23.85 ± 2.35 vs 10.94 ± 0.23),and induced a significant migratory(4.0-fold) and invasive(5.7-fold) effect.TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.CONCLUSION:These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.
文摘Objective: To study the effect of laparoscopic surgery and laparotomy on oxidative stress response and cell invasion in lesions after hysteromyomectomy. Methods: Patients with uterine fibroids who received surgical resection in our hospital between August 2014 and March 2017 were retrospectively analyzed and divided into laparoscopy group and laparotomy group according to different surgical procedures. Immediately after surgery and 24 h after surgery, the contents of oxidative stress response indexes in serum were measured;after surgical resection, the uterine fibroid lesion was collected to determine the expression of cell invasion indexes. Results: Serum MDA, NE, E, Cor and ACTH levels of laparoscopy group immediately after surgery and 24 h after surgery were significantly lower than those of laparotomy group while GSH-Px and T-SOD levels were significantly higher than those of laparotomy group;after surgical resection, CXCL12, CXCR4, MMP2, MMP7 and MMP9 mRNA expression in uterine fibroid lesions of laparoscopy group were significantly lower than those of laparotomy group while TIMP1, TIMP2, RECK and E-cadherin mRNA expression were significantly higher than those of laparotomy group. Conclusion: Laparoscopic surgery can reduce the oxidative stress response after hysteromyoma and inhibit the invasive growth of cells in the lesion.
基金Supported by National Natural Science Foundation of China,No.81071965
文摘AIM:To profile expression of micro RNAs(mi RNAs)in gastric cancer cells and investigate the effect of mi R-374b-5p on gastric cancer cell invasion and metastasis.METHODS:An mi RNA microarray assay was performed to identify mi RNAs differentially expressed in gastric cancer cell lines(MGC-803 and SGC-7901)compared with a normal gastric epithelial cell line.Upregulation of mi R-374b-5p was newly identified and confirmed via quantitative real-time reverse transcriptionPCR(q RT-PCR).MGC-803 cells were transfected with a synthesized anti-mi R-374b-5p sequence or a control vector using Lipofectamine reagent,or treated with transfection reagent alone or phosphate-buffered saline as controls.Rate of transfection was verified after 48 h by q RT-PCR.Cells were then subjected to transwell migration,wound scratch and cell counting kit-8 assays.A bioinformatic analysis to identify mi R-374b-5p target genes was performed using mi Randa,Pic Tar and Target Scan software.A dual luciferase reporter assay was performed to evaluate the influence of mi R-374b-5p on target gene activation,and q RT-PCR and Western blot were used to evaluate the levels of target m RNA and protein following transfection with mi R-374b-5p antisense oligonucleotides.RESULTS:The microarray profiling revealed downregulation of 14(fold change<0.667;P<0.05)and upregulation of 12(fold change>1.50;P<0.05)mi RNAs in MGC-803 and SGC-7901 cells compared with GES-1 controls.The upregulation of mi R-374b-5p(fold change=1.75 and 1.64 in MGC-803 and SGC-7901,respectively;P<0.05)was confirmed by q RT-PCR.Compared with the control groups,the restoration of mi R-374b-5p expression with anti-mi R-374b-5p significantly suppressed the metastasis,invasion and proliferation of MGC-803 cells.The bioinformatic analysis predicted that the 3’untranslated region(UTR)of reversion-inducing cysteine-rich protein with Kazal motif(RECK)contains three mi R-374b-5p target sequences.RECK was verified as a target gene in a dual luciferase reporter assay showing that activation of RECK 3’UTR-pmir GLO was increased by co-transfection with mi R-374b-5p.Finally,transfection of mi R-374b-5p antisense oligonucleotides increased m RNA and protein levels of RECK in MGC-803cells(P<0.05).CONCLUSION:These findings indicate that upregulation of mi R-374b-5p contributes to gastric cancer cell metastasis and invasion through inhibition of RECK expression.
基金Supported by The Affiliated First People’s Hospital, Shanghai Jiao Tong University and the Board of Education Fund for Scientific Research of Shanghai, China, No. 06BE067
文摘AIM: To investigate RNA interference targeting signal transducer and activator of transcription-3 (STAT3) on invasion of human pancreatic cancer cells. METHODS: We constructed three plasmids of RNA interference targeting the STAT3 gene. After LV (lentivirus)-STAT3siRNA (STAT3 small interfering RNA) the vector was transfected into the human pancreatic cell line, SW1990 and cell proliferation was measured by the MTT assay. Flow cytometry was used to assess cell cycle. Vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) mRNA and protein expression were examined by quantitative PCR and western blotting, respectively. The invasion ability of SW1990 cells was determined by cell invasion assay. RESULTS: We successfully constructed the LVSTAT3siRNA lentivirus vector and proved that it can suppress expression of STAT3 gene in SW1990 cells. RNA interference of STAT3 by the LV-STAT3siRNA construct signifi cantly inhibited the growth of SW1990 cells, in addition to signifi cantly decreasing both VEGF and MMP-2 mRNA and protein expression. Moreover, suppression of STAT3 by LV-STAT3siRNA decreased the invasion ability of SW1990 cells.CONCLUSION: The STAT3 signaling pathway may provide a novel therapeutic target for the treatment of pancreatic cancer since it inhibits the invasion ability of pancreatic cancer cells.
基金Projects of Shandong Provincial Natural Science Foundation(Grant No.81470709).
文摘Objective:To study the correlation of c-fos and p16 expression in oral squamous cell carcinoma with cell cycle and cell invasion.Methods: Patients with oral squamous cell carcinoma who underwent surgical resection in Zaozhuang Mining Group Central Hospital between June 2014 and March 2017 were selected as OSCC group of the research, and patients who received impacted tooth extraction and provided normal gingival mucosa tissue in the Zaozhuang Mining Group Central Hospital during the same period were selected as the control group. The lesion tissue and normal tissue were obtained from OSCC group and control group respectively to determine the expression of c-fos, p16, cell cycle molecules and cell invasion molecules.Results: c-fos mRNA expression in lesion tissue of OSCC group was significantly higher than that of control group, p16 mRNA expression was significantly lower than that of control group, and the c-fos mRNA expression was negatively correlated with p16 mRNA expression;Chk1 and Chk2 mRNA expression in lesion tissue of OSCC group were significantly lower than those of control group whereas NEK2, CyclinD1, MALAT1, Periostin,β-catenin and Vimentin mRNA expression were significantly higher than those of control group;Chk1 and Chk2 mRNA expression in OSCC lesion tissue with high c-fos expression were significantly lower than those in OSCC lesion tissue with low c-fos expression whereas NEK2, CyclinD1, MALAT1, Periostin,β-catenin and Vimentin mRNA expression were significantly higher than those in OSCC lesion tissue with low c-fos expression.Conclusion:The high expression of c-fos and the low expression of p16 in oral squamous cell carcinoma can accelerate cell cycle and promote cell invasion.
文摘Objective:To study the correlation of miR-106b in nasopharyngeal carcinoma with cell cycle progression and cell invasion.Methods: Patients who were diagnosed with nasopharyngeal carcinoma by pathological biopsy in Shenmu Hospital between March 2013 and February 2018 were included in the study, and the nasopharyngeal carcinoma tissue and tissue adjacent to nasopharyngeal carcinoma obtained from biopsy were collected, miRNA was extracted to determine miR-106b expression, and RNA was extracted to determine the mRNA expression of cell cycle-related genes and cell invasion-related genes.Results: miR-106b expression as well as FOXC1, CyclinD1, CyclinE, TRIM27, uPAR, PARP1 and MTA1 mRNA expression in nasopharyngeal carcinoma tissues were higher than those in adjacent tissues, CYB5R2, p16, IRF5 and E-cadherin mRNA expression were significantly lower than those in adjacent tissues whereas;CYB5R2, p16, IRF5 and E-cadherin mRNA expression in nasopharyngeal carcinoma tissues with high miR-106b expression were lower than those in nasopharyngeal carcinoma tissues with low miR-106b expression whereas FOXC1, CyclinD1, CyclinE, TRIM27, uPAR, PARP1 and MTA1 mRNA expression were higher than those in nasopharyngeal carcinoma tissues with low miR-106b expression.Conclusion:The lowly expressed miR-106b in nasopharyngeal carcinoma can promote the cell cycle progression and cell invasion in the lesion.
文摘Objective: To study the correlation of S100A13 and FOXA1 expression with cell cycle and cell invasion in fine needle aspiration thyroid carcinoma tissue. Methods: Patients who received ultrasound-guided thyroid nodule fine needle aspiration in Haiyang People's Hospital between April 2015 and February 2017 were selected, and the tissues were divided into malignant thyroid tissue and benign thyroid nodules according to the pathological results after biopsy. The expression of S100A13, FOXA1, cell cycle molecules and cell invasion molecules were measured. Results: S100A13, FOXA1, CDK2, CyclinD1, MCM2, MCM7, SKP2, CLOCK, STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue were significantly higher than those in benign thyroid nodule;CDK2, CyclinD1, MCM2, MCM7, SKP2 and CLOCK mRNA expression in thyroid carcinoma tissue with high FOXA1 expression were significantly higher than those in thyroid carcinoma tissue with low FOXA1 expression;STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue with high S100A13 expression were significantly higher than those in thyroid cancer tissue with low S100A13 expression. Conclusions: High expression of S100A13 and FOXA1 in thyroid carcinoma can promote cell invasion and cell cycle progression.
文摘Objective:To study the effects of high-intensity focused ultrasound (HIFU) combined with interventional chemoembolization on the advanced cervical cancer lesion growth and cell invasion.Methods:Patients with stage IIB-IVA cervical cancer treated in Suining Hospital of TCM between May 2014 and October 2016 were selected and randomly divided into two groups, HIFU group received HIFU combined with interventional chemoembolization, and the control group accepted interventional chemoembolization. The levels of tumor markers in serum as well as the expression of tumor suppressor genes and invasion genes in tumor lesions were determined before and after treatment.Results: 2 weeks, 3 weeks and 4 weeks after treatment, serum TK-1, SCC and CA125 levels of both groups were lower than those before treatment, serum TK-1, SCC and CA125 levels of HIFU group 2 weeks after treatment were not different from those of control group, and serum TK-1, SCC and CA125 levels of HIFU group 3 weeks and 4 weeks after treatment were significantly lower than those of control group;4 weeks after treatment, AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of both groups were higher than those before treatment while FAK, KGFR and MMP9 mRNA expression were lower than those before treatment, and AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of HIFU group were higher than those of control group while FAK, KGFR and MMP9 mRNA expression were lower than those of control group.Conclusion: HIFU combined with interventional chemoembolization can be more effective in suppressing the advanced cervical cancer lesion growth and cell invasion than interventional chemoembolization alone.
基金The financial supports provided,in whole or in part,by the National Natural Science Foundation of China(11272083)the New Century Excellent Talents Program in Chinese Universities(NCET09-0263)
文摘Introduction Hematogenous metastasis is the mainly leading cause of death in breast carcinoma patients.A better understanding of the underlying molecular and cellular mechanisms is crucial for the development of effective treatment for metastatic breast cancer<sup>[</sup>1].It has been well established that cell adhesion and invasion is mediated by a variety of transmembrane proteins,including integrins,cadherins,selectins,and intercellular adhesion molecules.Among these adhesion molecules,the integrins and their downstream signaling pathways have been extensively studied<sup>[2]</sup>.On the other hand,the specific events determining tumor cell interactions with endo-
文摘Ovarian cancer(OC)is a major cause of cancer-related deaths among gynaecologicalmalignancies.Emerging studies suggest that the long non-coding RNA(lncRNA)may be the potential biomarker for the diagnosis and prognosis of the cancer.The current study was carried out to investigate the role of lncRNA CCHE1 silencing in OC cell invasion and migration.Expression of lncRNA CCHE1 in normal ovarian cell Hose and OC cell lines HO 8910,A2780 and SKOV3 was detected.LncRNA were transfected with siRNA,and then the proliferation of cells was detected by using MTT assay.Cell invasion and migration was measured by using Transwell assay and scratch test,respectively.The protein levels of E-cadherin,N-cadherin,ERK,p38-MAPK and the phosphorylation of ERK and p38-MAPK in cells after siRNA transfection were detected by using Western blot analysis.Consequently,lncRNA CCHE1 expression was highly expressed in OC cell lines,especially in SKOV3 cells.siRNA1,siRNA2 and siRNA3 all decreased.lncRNA CCHE1 expression in SKOV3 cells and siRNA2 showed the best silencing efficacy.Silencing of lncRNA CCHE1 decreased proliferation,invasion and migration,and reduced the protein levels of N-cadherin,ERK,p38-MAPK and the phosphorylation of ERK and p38-MAPK,while reducing the protein level of E-cadherin in SKOV3 cells.Collectively,our study proved that the silencing of lncRNA CCHE1 could inhibit SKOV3 cell invasion and migration via inactivating the p38-MAPK signaling pathway.
文摘Objective:To study the relationship between osteopontin expression and cell invasion as well as angiogenesis in preeclampsia placenta tissue.Methods:A total of 56 patients diagnosed with preeclampsia in our hospital between June 2014 and March 2016 were selected as preeclampsia group (PE group) and 80 normal subjects receiving antenatal examination and giving birth during the same period were selected as control group. After labor, the placenta tissue was collected, immunohistochemical staining kits were used to detect OPN expression and enzyme-linked immunosorbent assay kits were used to determine cell invasion and angiogenesis molecule expression.Results:Positive OPN rate in placenta tissue of PE group was significantly lower than that of control group;cell invasion molecules LAMA4, uPA, Rho and ROCK as well as angiogenesis molecules VEGF, PIGF and AGF protein expression in placenta tissue of PE group were significantly lower than those of control group, and cell invasion molecules LAMA4, uPA, Rho and ROCK as well as angiogenesis molecules VEGF, PIGF and AGF protein expression in OPN-positive placenta tissue were significantly higher than those in OPN-negative placenta tissue.Conclusion:Lowly expressed OPN in preeclampsia placenta tissue will inhibit the expression of cell invasion molecules and angiogenesis molecules so as to inhibit sertoli cell invasion and the angiogenesis in placenta tissue.
文摘Objective: To study the effect of mesoderm-specific transcript (MEST) on the angiogenesis and trophoblast cell invasion in the preeclampsia placenta. Methods: The puerperae who were diagnosed with preeclampsia and gave birth in our hospital between June 2014 and February 2017 were selected as the PE group, and the healthy puerperae who gave birth in our hospital during the same period were selected as the control group. After delivery, the placental tissue was collected, fluorescence quantitative PCR method was used to determine MEST mRNA expression, and enzyme-linked immunosorbent assay was used to determine the protein levels of angiogenesis molecules and cell invasion molecules. Results: MEST, Notch1, DLL4, VEGF, Rho, ROCK, LAMA4, Notch2 and Notch3 protein levels in placental tissue of PE group were obviously lower than those of control group whereas DNC, sFlt-1 and sEng protein levels were obviously higher than those of control group;Notch1, DLL4, VEGF, Rho, ROCK, LAMA4, Notch2 and Notch3 protein levels in PE placenta tissue with lower MEST expression were greatly lower than those in PE placenta tissue with higher MEST expression whereas DNC, sFlt-1 and sEng protein levels were greatly higher than those in PE placenta tissue with higher MEST expression. Conclusion: The low expression of MEST in preeclampsia placenta can inhibit the angiogenesis and trophoblast cell invasion.
