The prognosis for patients who are diagnosed with advanced stage hepatocellular carcinoma(HCC)is poor because there are few treatment options.Recent research has focused on the identification of novel molecular entiti...The prognosis for patients who are diagnosed with advanced stage hepatocellular carcinoma(HCC)is poor because there are few treatment options.Recent research has focused on the identification of novel molecular entities that can be targeted to inhibit oncogenic signals that are involved in the carcinogenesis,proliferation and progression of HCC.Among all of the pathways that are involved in the development of HCC,Hedgehog(HH)signalling has demonstrated a substantial role in hepatocarcinogenesis and HCC progression.HH plays a physiological role in embryogenesis,through the induction of the differentiation of hepatocytes from endodermal progenitors.The re-activation of the HH pathway in chronic damaged liver is a mechanism of fibrotic degeneration and is implicated in various stages of HCC development.HH activation sustains the subpopulation of immature liver epithelial cells that are involved in the pathogenesis of cirrhosis and HCC,and HH itself is a mediator of the alcohol-derived malignant transformation of liver cells.High levels of expression of HH protein markers in liver tumour tissues are correlated with aggressive histological and biological features and a poor clinical outcome.In vitro and in vivo inhibition models of the HH pathway confirm that HH is essential in maintaining tumour growth,metastasis and a mesenchymal phenotype.展开更多
AIM:To investigate the effect of hyperthermia on hy-poxia-induced epithelial-mesenchymal transition (EMT) in HepG2 hepatocellular carcinoma (HCC) cells, and its mechanism. METHODS:Cells were treated with hyperthermia ...AIM:To investigate the effect of hyperthermia on hy-poxia-induced epithelial-mesenchymal transition (EMT) in HepG2 hepatocellular carcinoma (HCC) cells, and its mechanism. METHODS:Cells were treated with hyperthermia at 43 ℃ for 0.5 h, followed by incubation under hypoxic or normoxic conditions for 72 h. Cell morphology was observed. Expressions of E-cadherin and vimentin were determined by immunofluorescence assay or Western blot. The protein and mRNA expressions of Snail were also determined by Western blot and reverse transcrip-tion-polymerase chain reaction. Cell migratory capacity was evaluated. RESULTS:Hypoxia induced EMT in HepG2 cells, which was evidenced by morphological, molecular and func-tional changes, including the formation of a spindle shape and the loss of cell contact. The expression of E-cadherin was decreased but the expression of vimentin was increased; also, the migratory capability was increased by 2.2 ± 0.20-fold as compared with normoxia. However, those effects were inhibited by hyperthermia pretreatment. Furthermore, protein synthesis and mRNA expression of Snail in the cells were enhanced by hy-poxia as compared with normoxia, and also significantly inhibited by hyperthermia pretreatment. CONCLUSION:Hyperthermia may inhibit hypoxia-induced EMT in HepG2 HCC cells, and the mechanism may involve inhibition of induced expression of Snail.展开更多
Pancreatic cancer has one of the worst prognoses among all cancers due to the late manifestation of identifiable symptoms and high resistance to chemo- and radiation therapies. In recent years, a cancer development ph...Pancreatic cancer has one of the worst prognoses among all cancers due to the late manifestation of identifiable symptoms and high resistance to chemo- and radiation therapies. In recent years, a cancer development phase termed epithelial-mesenchymal transition(EMT) has gained increasing research focus. The process is implicated in tumour metastasis, and emerging evidence suggests EMT also contributes or induces chemoresistance in several cancers. Nevertheless, the applicability of therapeutic targeting of EMT faces many challenges. In this mini-review, we summarise the evidence supporting the role of EMT in pancreatic cancer progression, focusing particularly on its association with chemoresistance.展开更多
AIM: To study the effect of discoidin I-like domaincontaining protein 3(EDIL3) depletion on the proliferation and epithelial-mesenchymal transition(EMT) in human lens epithelial cells(LECs). METHODS: RNA interference ...AIM: To study the effect of discoidin I-like domaincontaining protein 3(EDIL3) depletion on the proliferation and epithelial-mesenchymal transition(EMT) in human lens epithelial cells(LECs). METHODS: RNA interference was used to inhibit the expression of EDIL3 in human LECs in vitro. The morphology of cells was observed using an inverted microscope. Cell proliferation was assessed using Ed U kit. Cell migration was investigated using Transwell chamber and EMT of LECs was assessed using confocal microscope and Western blotting. The transforming growth factor β(TGFβ) pathway was investigated using Western blotting. RESULTS: The data showed that silencing EDIL3 expression changed LECs morphology and suppressed LECs proliferation(P<0.05) and migration(P<0.01). Furthermore, the result of Western blotting showed that EDIL3 depletion reduced the expression of α-smooth muscle actin(α-SMA)(P<0.001) and vimentin(P<0.01), while increased the expression of E-cadherin(P<0.001). EDIL3 depletion could suppress the phosphorylation of Smad2(P<0.01) and Smad3(P<0.01) and the activation of exracellular signal regulated kinase(ERK)(P<0.05). CONCLUSION: The findings indicate that EDIL3 might participate in the proliferation and EMT in LECs via TGFβ pathway and may be a potential therapeutic target for the treatment of posterior capsule opacification.展开更多
Epithelial-mesenchymal transition(EMT) has been linked with aggressive tumor biology and therapy resistance. It plays central role not only in the generation of cancer stem cells(CSCs) but also direct them across the ...Epithelial-mesenchymal transition(EMT) has been linked with aggressive tumor biology and therapy resistance. It plays central role not only in the generation of cancer stem cells(CSCs) but also direct them across the multiple organ systems to promote tumor recurrence and metastasis. CSCs are reported to express stem cell genes as well as specific cell surfacemarkers and allow aberrant differentiation of progenies.It facilitates cancer cells to leave primary tumor, acquire migratory characteristics, grow into new environment and develop radio-chemo-resistance. Based on the current information, present review discusses and summarizes the recent advancements on the molecular mechanisms that derive epithelial plasticity and its major role in generating a subset of tumor cells with stemness properties and pathophysiological spread of tumor. This paper further highlights the critical need to examine the regulation of EMT and CSC pathways in identifying the novel probable therapeutic targets.These improved therapeutic strategies based on the co-administration of inhibitors of EMT, CSCs as well as differentiated tumor cells may provide improved antineoplastic response with no tumor relapse.展开更多
AIM: To investigate the expression of transcription factors Slug in human lens epithelial cells(HLECs)undergoing epithelial-mesenchymal transition(EMT)induced by connective tissue growth factor(CTGF).·METHODS: HL...AIM: To investigate the expression of transcription factors Slug in human lens epithelial cells(HLECs)undergoing epithelial-mesenchymal transition(EMT)induced by connective tissue growth factor(CTGF).·METHODS: HLECs were treated with CTGF of different concentrations(20, 50 and 100 ng/m L) or without CTGF(control) for 24 h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin(α-SMA) were further determined by Western blot analysis.·RESULTS: HLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64 ±0.11, 1.96 ±0.03,3.12 ±0.10, and 4.08 ±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and100 ng/m L(F =443.86, P <0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA(0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F =449.85, P <0.01) and down-expression of E-cadherin(2.50 ±0.11,1.79±0.26, 1.05±0.14, 0.63±0.08; F =101.55, P <0.01).·CONCLUSION: Transcription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.展开更多
AIM To elucidate the effect of expression of doublecortin and Ca M kinase-like-1(DCLK1) in patients with pancreatic ductal adenocarcinoma(PDAC). METHODS Tumor specimens were obtained from 136 patients with pancreatic ...AIM To elucidate the effect of expression of doublecortin and Ca M kinase-like-1(DCLK1) in patients with pancreatic ductal adenocarcinoma(PDAC). METHODS Tumor specimens were obtained from 136 patients with pancreatic cancer who had undergone resection without preoperative therapy between January 2000 and December 2013 at the Department of Surgical Oncology, Osaka City University. The resected specimens were analyzed for associations with clinicopathological data, including DCLK1 expression, epithelial mesenchymal transition(EMT) marker expression, and cancer stem cell(CSC) marker expression. Univariate and multivariate survival analyses were performed and we assessed the association between DCLK1 expression and clinicopathological factors, including the EMT marker and CSC marker.RESULTS In total, 48.5%(66/136) of the pancreatic cancer samples were positive for DCLK1. Patients with DCLK1-positive tumors had significantly shorter survival times than those with DCLK1-negative tumors(median, 18.7 mo vs 49.5 mo, respectively; P < 0.0001). Positive DCLK1 expression correlated with histological grade(P = 0.0290), preoperative CA19-9 level(P = 0.0060), epithelial cell adhesion molecule(Ep CAM) expression(P = 0.0235), and the triple-positive expression of CD44/CD24/Ep CAM(P = 0.0139). On univariate survival analysis, five factors were significantly associated with worse overall survival: histological grade of G2 to G4(P = 0.0091), high preoperative serum SPan-1 level(P = 0.0034), R1/2(P < 0.0001), positive expression of DCLK1(P < 0.0001) or CD44(P = 0.0245). On multivariate survival analysis, R1/2 [odds ratio(OR) = 2.019, 95% confidence interval(CI): 1.380-2.933; P = 0.0004] and positive DCLK1 expression(OR = 1.848, 95%CI: 1.2854-2.661; P = 0.0009) were independent prognostic factors. CONCLUSION DCLK1 expression was found to be an independent prognostic factor and it may play a crucial prognostic role by promoting acquisition of stemness.展开更多
AIM:To evaluate the effect of exogenous recombinant human bone morphogenic protein-7(rhBMP-7)on transforming growth factor-β(TGF-β)-induced epithelial mesenchymal cell transition(EMT)and assessed its antifibrotic ef...AIM:To evaluate the effect of exogenous recombinant human bone morphogenic protein-7(rhBMP-7)on transforming growth factor-β(TGF-β)-induced epithelial mesenchymal cell transition(EMT)and assessed its antifibrotic effect via topical application.METHODS:The cytotoxic effect of rhBMP-7 was evaluated and the EMT of human corneal epithelial cells(HECEs)was induced by TGF-β. HECEs were then cultured in the presence of rhBMP-7 and/or hyaluronic acid(HA). EMT markers,fibronectin,E-cadherin,α-smooth muscle actin(α-SMA),and matrix metaloproteinase-9(MMP-9),were evaluated. The level of corneal fibrosis and the reepithelization rate were evaluated using a rabbit keratectomy model. Expression of α-SMA in keratocytes were quantified following treatment with different concentrations of rhBMP-7.RESULTS:Treatment with rhBMP-7 attenuated TGF-β-induced EMT in HECEs. It significantly attenuated fibronectin secretion(31.6%; P<0.05),the α-SMA protein level(72.2%; P<0.01),and MMP-9 expression(23.6%,P<0.05)in HECEs compared with cells grown in the presence of TGF-β alone. E-cadherin expression was significantly enhanced(289.7%; P<0.01)in the presence of rhBMP-7. Topical application of rhBMP-7 combined with 0.1% HA significantly reduced the amount of α-SMA^+ cells by 43.18%(P<0.05)at a concentration of 2.5 μg/mL and by 47.73%(P<0.05)at 25 μg/mL,compared with the control group,without disturbing corneal reepithelization.CONCLUSION:rhBMP-7 attenuates TGF-β-induced EMT in vitro,and topical application of rhBMP-7 reduces keratocyte myodifferentiation during the early wound healing stages in vivo without hindering reepithelization. Topical rhBMP-7 application as biological eye drops seems to be feasible in diseases involving TGF-β-related corneal fibrosis with corneal reepithelization disorders.展开更多
AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition(EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery(FLACS).METHODS: Sixty cataract patients w...AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition(EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery(FLACS).METHODS: Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly: FLACS1 group(cataract surgery by FLACS with LenS x), FLACS2 group(cataract surgery by FLACS with LensA R) and manual group(cataract surgery by phacoemulsification). Patients in two FLACS groups performed anterior capsulotomy by Len Sx or Lens AR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis(CCC) manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery.RESULTS: The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC(P<0.05). Lens epithelium cells apoptosis were correlatedwith femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT.CONCLUSION: Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.展开更多
AIM To explore the functional role of cullin 4A(CUL4A), a core subunit of E3 ubiquitin ligase, in perihilar cholangiocarcinoma(PHCC).METHODS The expression of CUL4 A in PHCC cell lines was evaluated by Western blot an...AIM To explore the functional role of cullin 4A(CUL4A), a core subunit of E3 ubiquitin ligase, in perihilar cholangiocarcinoma(PHCC).METHODS The expression of CUL4 A in PHCC cell lines was evaluated by Western blot and quantitative reverse transcription-polymerase chain reaction. Immunohistochemistry(IHC) was adopted to investigate the relationship between CUL4 A expression and clinicopathological characteristics of PHCC. Univariate analysis and multivariate regression analysis were performed to analyze the risk factors related to overall survival(OS) and progression-free survival(PFS) of PHCC patients. Wound healing, Transwell and Matrigel assays were utilized to explore the function of CUL4 A in PHCC metastasis. Furthermore, expression of epithelial to mesenchymal transition(EMT) markers was verified in cells with CUL4 A knockdown or overexpression. The relationship between CUL4 A expression and E-cadherin expression was also analyzed by IHC assay. Finally, the role of ZEB1 in regulating CUL4 A mediated PHCC was detected by IHC, Western blot, Transwell and Matrigel assays.RESULTS CUL4 A overexpression was detected in PHCC cell lines and clinical specimens. Clinicopathological analysis revealed a close correlation between CUL4 A overexpression and tumour differentiation, T, N and TNM stages in PHCC. Kaplan-Meier analysis revealed that high CUL4 A expression was correlated with poor OS and PFS of PHCC patients. Univariate analysis identified the following four parameters as risk factors related to OS rate of PHCC: T, N, TNM stages and high CUL4 A expression; as well as three related to PFS: N stage, TNM stage and high CUL4 A expression. Further multivariate logistic regression analysis identified high CUL4 A expression as the only independent prognostic factor for PHCC. Moreover, CUL4 A silencing in PHCC cell lines dramatically inhibited metastasis and the EMT. Conversely, CUL4 A overexpression promoted these processes. Mechanistically, ZEB1 was discovered to regulate the function of CUL4 A in promoting the EMT and metastasis.CONCLUSION CUL4 A is an independent prognostic factor for PHCC, and it can promote the EMT by regulating ZEB1 expression. CUL4 A may be a potential therapeutic target for PHCC.展开更多
文摘The prognosis for patients who are diagnosed with advanced stage hepatocellular carcinoma(HCC)is poor because there are few treatment options.Recent research has focused on the identification of novel molecular entities that can be targeted to inhibit oncogenic signals that are involved in the carcinogenesis,proliferation and progression of HCC.Among all of the pathways that are involved in the development of HCC,Hedgehog(HH)signalling has demonstrated a substantial role in hepatocarcinogenesis and HCC progression.HH plays a physiological role in embryogenesis,through the induction of the differentiation of hepatocytes from endodermal progenitors.The re-activation of the HH pathway in chronic damaged liver is a mechanism of fibrotic degeneration and is implicated in various stages of HCC development.HH activation sustains the subpopulation of immature liver epithelial cells that are involved in the pathogenesis of cirrhosis and HCC,and HH itself is a mediator of the alcohol-derived malignant transformation of liver cells.High levels of expression of HH protein markers in liver tumour tissues are correlated with aggressive histological and biological features and a poor clinical outcome.In vitro and in vivo inhibition models of the HH pathway confirm that HH is essential in maintaining tumour growth,metastasis and a mesenchymal phenotype.
基金Supported by Medical Science and Technology Innovation Foundation of Nanjing Military Command of Chinese People’s Liberation Army, No. 11MA036Natural Science Foundation of China, No. 81000998
文摘AIM:To investigate the effect of hyperthermia on hy-poxia-induced epithelial-mesenchymal transition (EMT) in HepG2 hepatocellular carcinoma (HCC) cells, and its mechanism. METHODS:Cells were treated with hyperthermia at 43 ℃ for 0.5 h, followed by incubation under hypoxic or normoxic conditions for 72 h. Cell morphology was observed. Expressions of E-cadherin and vimentin were determined by immunofluorescence assay or Western blot. The protein and mRNA expressions of Snail were also determined by Western blot and reverse transcrip-tion-polymerase chain reaction. Cell migratory capacity was evaluated. RESULTS:Hypoxia induced EMT in HepG2 cells, which was evidenced by morphological, molecular and func-tional changes, including the formation of a spindle shape and the loss of cell contact. The expression of E-cadherin was decreased but the expression of vimentin was increased; also, the migratory capability was increased by 2.2 ± 0.20-fold as compared with normoxia. However, those effects were inhibited by hyperthermia pretreatment. Furthermore, protein synthesis and mRNA expression of Snail in the cells were enhanced by hy-poxia as compared with normoxia, and also significantly inhibited by hyperthermia pretreatment. CONCLUSION:Hyperthermia may inhibit hypoxia-induced EMT in HepG2 HCC cells, and the mechanism may involve inhibition of induced expression of Snail.
文摘Pancreatic cancer has one of the worst prognoses among all cancers due to the late manifestation of identifiable symptoms and high resistance to chemo- and radiation therapies. In recent years, a cancer development phase termed epithelial-mesenchymal transition(EMT) has gained increasing research focus. The process is implicated in tumour metastasis, and emerging evidence suggests EMT also contributes or induces chemoresistance in several cancers. Nevertheless, the applicability of therapeutic targeting of EMT faces many challenges. In this mini-review, we summarise the evidence supporting the role of EMT in pancreatic cancer progression, focusing particularly on its association with chemoresistance.
基金Supported by the National Natural Science Foundation of China (No.81700839)Military logistics scientific research project (No.BWS12J030)+2 种基金Natural Science Foundation of Shanghai (No.15ZR1413200)Research Foundation for Youth of Second Military Medical University (No.2016QN13)Research Foundation for Youth of Changhai Hospital (No.CH201712)
文摘AIM: To study the effect of discoidin I-like domaincontaining protein 3(EDIL3) depletion on the proliferation and epithelial-mesenchymal transition(EMT) in human lens epithelial cells(LECs). METHODS: RNA interference was used to inhibit the expression of EDIL3 in human LECs in vitro. The morphology of cells was observed using an inverted microscope. Cell proliferation was assessed using Ed U kit. Cell migration was investigated using Transwell chamber and EMT of LECs was assessed using confocal microscope and Western blotting. The transforming growth factor β(TGFβ) pathway was investigated using Western blotting. RESULTS: The data showed that silencing EDIL3 expression changed LECs morphology and suppressed LECs proliferation(P<0.05) and migration(P<0.01). Furthermore, the result of Western blotting showed that EDIL3 depletion reduced the expression of α-smooth muscle actin(α-SMA)(P<0.001) and vimentin(P<0.01), while increased the expression of E-cadherin(P<0.001). EDIL3 depletion could suppress the phosphorylation of Smad2(P<0.01) and Smad3(P<0.01) and the activation of exracellular signal regulated kinase(ERK)(P<0.05). CONCLUSION: The findings indicate that EDIL3 might participate in the proliferation and EMT in LECs via TGFβ pathway and may be a potential therapeutic target for the treatment of posterior capsule opacification.
文摘Epithelial-mesenchymal transition(EMT) has been linked with aggressive tumor biology and therapy resistance. It plays central role not only in the generation of cancer stem cells(CSCs) but also direct them across the multiple organ systems to promote tumor recurrence and metastasis. CSCs are reported to express stem cell genes as well as specific cell surfacemarkers and allow aberrant differentiation of progenies.It facilitates cancer cells to leave primary tumor, acquire migratory characteristics, grow into new environment and develop radio-chemo-resistance. Based on the current information, present review discusses and summarizes the recent advancements on the molecular mechanisms that derive epithelial plasticity and its major role in generating a subset of tumor cells with stemness properties and pathophysiological spread of tumor. This paper further highlights the critical need to examine the regulation of EMT and CSC pathways in identifying the novel probable therapeutic targets.These improved therapeutic strategies based on the co-administration of inhibitors of EMT, CSCs as well as differentiated tumor cells may provide improved antineoplastic response with no tumor relapse.
基金Supported by National Natural Science Foundation of China(No.81470614,No.81460163,No.81300786)Fundamental Research Funds for the Central Universities(No.xjj2014146)+1 种基金Specialized Research Fund for the Doctoral Program of Higher Education(No.20133601120012)Key International Communication Project of Shaanxi province(No.2012KW-31)
文摘AIM: To investigate the expression of transcription factors Slug in human lens epithelial cells(HLECs)undergoing epithelial-mesenchymal transition(EMT)induced by connective tissue growth factor(CTGF).·METHODS: HLECs were treated with CTGF of different concentrations(20, 50 and 100 ng/m L) or without CTGF(control) for 24 h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin(α-SMA) were further determined by Western blot analysis.·RESULTS: HLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64 ±0.11, 1.96 ±0.03,3.12 ±0.10, and 4.08 ±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and100 ng/m L(F =443.86, P <0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA(0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F =449.85, P <0.01) and down-expression of E-cadherin(2.50 ±0.11,1.79±0.26, 1.05±0.14, 0.63±0.08; F =101.55, P <0.01).·CONCLUSION: Transcription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.
文摘AIM To elucidate the effect of expression of doublecortin and Ca M kinase-like-1(DCLK1) in patients with pancreatic ductal adenocarcinoma(PDAC). METHODS Tumor specimens were obtained from 136 patients with pancreatic cancer who had undergone resection without preoperative therapy between January 2000 and December 2013 at the Department of Surgical Oncology, Osaka City University. The resected specimens were analyzed for associations with clinicopathological data, including DCLK1 expression, epithelial mesenchymal transition(EMT) marker expression, and cancer stem cell(CSC) marker expression. Univariate and multivariate survival analyses were performed and we assessed the association between DCLK1 expression and clinicopathological factors, including the EMT marker and CSC marker.RESULTS In total, 48.5%(66/136) of the pancreatic cancer samples were positive for DCLK1. Patients with DCLK1-positive tumors had significantly shorter survival times than those with DCLK1-negative tumors(median, 18.7 mo vs 49.5 mo, respectively; P < 0.0001). Positive DCLK1 expression correlated with histological grade(P = 0.0290), preoperative CA19-9 level(P = 0.0060), epithelial cell adhesion molecule(Ep CAM) expression(P = 0.0235), and the triple-positive expression of CD44/CD24/Ep CAM(P = 0.0139). On univariate survival analysis, five factors were significantly associated with worse overall survival: histological grade of G2 to G4(P = 0.0091), high preoperative serum SPan-1 level(P = 0.0034), R1/2(P < 0.0001), positive expression of DCLK1(P < 0.0001) or CD44(P = 0.0245). On multivariate survival analysis, R1/2 [odds ratio(OR) = 2.019, 95% confidence interval(CI): 1.380-2.933; P = 0.0004] and positive DCLK1 expression(OR = 1.848, 95%CI: 1.2854-2.661; P = 0.0009) were independent prognostic factors. CONCLUSION DCLK1 expression was found to be an independent prognostic factor and it may play a crucial prognostic role by promoting acquisition of stemness.
基金Supported by the Soonchunhyang University Research Fund,the WPM project,Ministry of trade,industry&energy(No.10037842)the National Research Foundation of Korea(No.NRF-2016R1C1B2015622)Recombinant human BMP-7 protein was kindly provided by Cellumed Co.,Ltd
文摘AIM:To evaluate the effect of exogenous recombinant human bone morphogenic protein-7(rhBMP-7)on transforming growth factor-β(TGF-β)-induced epithelial mesenchymal cell transition(EMT)and assessed its antifibrotic effect via topical application.METHODS:The cytotoxic effect of rhBMP-7 was evaluated and the EMT of human corneal epithelial cells(HECEs)was induced by TGF-β. HECEs were then cultured in the presence of rhBMP-7 and/or hyaluronic acid(HA). EMT markers,fibronectin,E-cadherin,α-smooth muscle actin(α-SMA),and matrix metaloproteinase-9(MMP-9),were evaluated. The level of corneal fibrosis and the reepithelization rate were evaluated using a rabbit keratectomy model. Expression of α-SMA in keratocytes were quantified following treatment with different concentrations of rhBMP-7.RESULTS:Treatment with rhBMP-7 attenuated TGF-β-induced EMT in HECEs. It significantly attenuated fibronectin secretion(31.6%; P<0.05),the α-SMA protein level(72.2%; P<0.01),and MMP-9 expression(23.6%,P<0.05)in HECEs compared with cells grown in the presence of TGF-β alone. E-cadherin expression was significantly enhanced(289.7%; P<0.01)in the presence of rhBMP-7. Topical application of rhBMP-7 combined with 0.1% HA significantly reduced the amount of α-SMA^+ cells by 43.18%(P<0.05)at a concentration of 2.5 μg/mL and by 47.73%(P<0.05)at 25 μg/mL,compared with the control group,without disturbing corneal reepithelization.CONCLUSION:rhBMP-7 attenuates TGF-β-induced EMT in vitro,and topical application of rhBMP-7 reduces keratocyte myodifferentiation during the early wound healing stages in vivo without hindering reepithelization. Topical rhBMP-7 application as biological eye drops seems to be feasible in diseases involving TGF-β-related corneal fibrosis with corneal reepithelization disorders.
文摘AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition(EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery(FLACS).METHODS: Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly: FLACS1 group(cataract surgery by FLACS with LenS x), FLACS2 group(cataract surgery by FLACS with LensA R) and manual group(cataract surgery by phacoemulsification). Patients in two FLACS groups performed anterior capsulotomy by Len Sx or Lens AR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis(CCC) manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery.RESULTS: The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC(P<0.05). Lens epithelium cells apoptosis were correlatedwith femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT.CONCLUSION: Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.
文摘AIM To explore the functional role of cullin 4A(CUL4A), a core subunit of E3 ubiquitin ligase, in perihilar cholangiocarcinoma(PHCC).METHODS The expression of CUL4 A in PHCC cell lines was evaluated by Western blot and quantitative reverse transcription-polymerase chain reaction. Immunohistochemistry(IHC) was adopted to investigate the relationship between CUL4 A expression and clinicopathological characteristics of PHCC. Univariate analysis and multivariate regression analysis were performed to analyze the risk factors related to overall survival(OS) and progression-free survival(PFS) of PHCC patients. Wound healing, Transwell and Matrigel assays were utilized to explore the function of CUL4 A in PHCC metastasis. Furthermore, expression of epithelial to mesenchymal transition(EMT) markers was verified in cells with CUL4 A knockdown or overexpression. The relationship between CUL4 A expression and E-cadherin expression was also analyzed by IHC assay. Finally, the role of ZEB1 in regulating CUL4 A mediated PHCC was detected by IHC, Western blot, Transwell and Matrigel assays.RESULTS CUL4 A overexpression was detected in PHCC cell lines and clinical specimens. Clinicopathological analysis revealed a close correlation between CUL4 A overexpression and tumour differentiation, T, N and TNM stages in PHCC. Kaplan-Meier analysis revealed that high CUL4 A expression was correlated with poor OS and PFS of PHCC patients. Univariate analysis identified the following four parameters as risk factors related to OS rate of PHCC: T, N, TNM stages and high CUL4 A expression; as well as three related to PFS: N stage, TNM stage and high CUL4 A expression. Further multivariate logistic regression analysis identified high CUL4 A expression as the only independent prognostic factor for PHCC. Moreover, CUL4 A silencing in PHCC cell lines dramatically inhibited metastasis and the EMT. Conversely, CUL4 A overexpression promoted these processes. Mechanistically, ZEB1 was discovered to regulate the function of CUL4 A in promoting the EMT and metastasis.CONCLUSION CUL4 A is an independent prognostic factor for PHCC, and it can promote the EMT by regulating ZEB1 expression. CUL4 A may be a potential therapeutic target for PHCC.
