AIM To evaluate the clinical properties of three subpopulations of circulating tumor cells(CTCs) undergoing epithelial-mesenchymal transition(EMT) in pancreatic ductal adenocarcinoma(PDAC) patients.METHODS We identifi...AIM To evaluate the clinical properties of three subpopulations of circulating tumor cells(CTCs) undergoing epithelial-mesenchymal transition(EMT) in pancreatic ductal adenocarcinoma(PDAC) patients.METHODS We identified CTCs for expression of the epithelial cell marker cytokeratin or epithelial cell adhesion molecule(EpCAM)(E-CTC), the mesenchymal cell markers vimentin and twist(M-CTC), or both(E/M-CTC) using the CanPatrol system. Between July 2014 and July 2016, 107 patients with PDAC were enrolled for CTC evaluation. CTC enumeration and classification were correlated with patient clinicopathological features and outcomes.RESULTS CTCs were detected in 78.5% of PDAC patients. The number of total CTCs ranged from 0 to 26 across all 107 patients, with a median value of six. CTC status correlated with lymph node metastasis, TNM stage, distant metastasis, blood lymphocyte counts, and neutrophil-to-lymphocyte ratio(NLR). Kaplan-Meier survival analysis showed that patients with ≥ 6 total CTCs had significantly decreased overall survival and progression-free survival compared with patients with < 6 total CTCs. The presence of M-CTCs was positively correlated with TNM stage(P < 0.01) and distant metastasis(P < 0.01). Additionally, lymphocyte counts and NLR in patients without CTCs were significantly different from those in patients testing positive for each CTC subpopulation(P < 0.01).CONCLUSION Classifying CTCs by EMT markers helps to identify the more aggressive CTC subpopulations and provides useful evidence for determining a suitable clinical approach.展开更多
目的:探讨姜黄素(curcumin,Cur)抑制胰腺癌细胞上皮间质转化(epithelial-mesenchymal transition,EMT)及其影响胰腺癌侵袭转移的潜在机制.方法:常规培养3种不同分化特征的胰腺癌细胞株PANC-1、AsPC-1、BxPC-3细胞,采用Western blot法检...目的:探讨姜黄素(curcumin,Cur)抑制胰腺癌细胞上皮间质转化(epithelial-mesenchymal transition,EMT)及其影响胰腺癌侵袭转移的潜在机制.方法:常规培养3种不同分化特征的胰腺癌细胞株PANC-1、AsPC-1、BxPC-3细胞,采用Western blot法检测其EMT标志物E-cadherin、Vimentin蛋白的表达.以终浓度10 ng/mL的转化生长因子-β1(transforming growth factor beta1,TGF-β1)处理低分化PANC-1细胞24 h后,在倒置显微镜下观察其形态变化.分别以10 ng/mL TGF-β1、10μmol/L Cur+10 ng/mL TGF-β1、10μmol/L Cur处理PANC-1细胞,另设空白对照组,分别采用Real-time PCR及Western blot法检测各组E-cadherin、Vimentin表达变化.结果:Western blot结果显示,PANC-1、AsPC-1、BxPC-3细胞的E-cadherin蛋白表达依次增强(1.00±00、1.36±0.04、2.14±0.06,P<0.05),而Vimentin表达则依次减弱(1.00±0.00、0.60±0.05、0.49±0.04,P<0.05),这表明低分化的PANC-1细胞间质特性最强.TGF-β1刺激PANC-1细胞发生典型的EMT形态变化.Real-time PCR和Western b l o t结果均显示,PA N C-1细胞经姜黄素及T G F-β1处理后,与对照组相比,T G F-β1组E-cadherin mRNA及蛋白表达均明显下调(分别为0.67±0.05、0.47±0.05,P<0.05 vs对照组),Vimentin基因mRNA及蛋白则明显上调(分别为2.38±0.14、1.43±0.07,P<0.05 vs对照组),说明TGF-β1促进PANC-1细胞EMT过程.而与TGF-β1组相比,Cur+TGF-β1及Cur组的E-cadherin表达具有上调趋势(mRNA分别为0.98±0.06、1.34±0.08,P<0.05 vs TGF-β1组;蛋白为0.32±0.04、0.68±0.06,P<0.05 vs TGF-β1组),Vimentin却下调(mRNA分别为0.63±0.08、0.99±0.07,P<0.05 vs TGF-β1组;蛋白分别为1.01±0.14、0.57±0.06,P<0.05vs TGF-β1组),其蛋白与mRNA表达结果基本一致.这表明姜黄素具有阻断TGF-β1诱导的PANC-1细胞EMT效应.结论:姜黄素能够阻断TGF-β1诱导的PANC-1细胞的EMT过程,从而抑制其侵袭转移.展开更多
Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to ...Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to be involved in the development of adenomyosis.We aimed to investigate whether adenomyosis-derived extracellular vesicles(AMEVs)are able to induce an EMT process in endometrial epithelial cells.In this study,AMEVs were isolated from patients with adenomyosis and characterized by transmission electron microscopy,Western blot,and nanoparticle tracking.Primary endometrial epithelial cells(EECs)were derived from normal endometrium tissues from patients with leiomyoma and co-cultured with AMEVs in vitro.AMEV uptake was examined by fluorescence confocal microscopy.The invasion of EECs was confirmed by Transwell assay.Immunohistochemistry,Western blot,and qRT-PCR were performed on EECs to illustrate the expression levels of cytokeratin 19,E-cadherin,vimentin,and zinc finger E-boxbinding homeobox 1(ZEB1).The results indicated that the cellular fluorescence intensity gradually increased after 48 h of co-culture,but decreased after 72 h.After co-culturing with AMEVs for 72 h,EECs expressed significantly lower levels of cytokeratin 19 and E-cadherin,and significantly higher levels of vimentin and ZEB1.Together these results demonstrated that AMEVs induce an EMT process and enhance the invasion of EECs.These changes may contribute to the pathogenesis and progression of adenomyosis.展开更多
Gastric cancer(GC) is the fifth most common malignancy in the world. The major cause of GC is chronic infection with Helicobacter pylori(H. pylori). Infection with H. pylori leads to an active inflammatory microenviro...Gastric cancer(GC) is the fifth most common malignancy in the world. The major cause of GC is chronic infection with Helicobacter pylori(H. pylori). Infection with H. pylori leads to an active inflammatory microenvironment that is maintained by immune cells such as T cells, macrophages, natural killer cells, among other cells. Immune cell dysfunction allows the initiation and accumulation of mutations in GC cells, inducing aberrant proliferation and protection from apoptosis. Meanwhile, immune cells can secrete certain signals, including cytokines, and chemokines, to alter intracellular signaling pathways in GC cells. Thus, GC cells obtain the ability to metastasize to lymph nodes by undergoing the epithelial-mesenchymal transition(EMT), whereby epithelial cells lose their epithelial attributes and acquire a mesenchymal cell phenotype. Metastasis is a leading cause of death for GC patients, and the involved mechanisms are still under investigation. In this review, we summarize the current research on how the inflammatory environment affects GC initiation and metastasis via EMT.展开更多
AIM: To investigate the mechanism by which mi R-19 a is up-regulated in gastric cancer(GC), which plays an oncogenic role.METHODS: In the present study, we investigated the role of mi R-19 a in gastric tissues as well...AIM: To investigate the mechanism by which mi R-19 a is up-regulated in gastric cancer(GC), which plays an oncogenic role.METHODS: In the present study, we investigated the role of mi R-19 a in gastric tissues as well as two GC cell lines. In vivo, we detected the basal expression level of mi R-19 a using real-time reverse transcription-PCR(RTPCR), and the relevance between expression of mi R-19 a and clinicopathological information was analyzed.In vitro, mi R-19 a was ectopically expressed using overexpression and knock-down strategies.RESULTS: Overexpression of mi R-19 a was significantly associated with metastasis of GC and inferior overall prognosis. However, no significant correlation was f o u n d b e t w e e n m i R- 1 9 a e x p r e s s i o n a n d o t h e r characteristics such as age, gender, tobacco, alcohol or tumor size. Cell proliferation, migration and invasion assays showed that overexpression of mi R-19 a promoted the proliferation, migration and invasion, and that overexpression of mi R-19 a promoted the epithelialmesenchymal transition through activating the PI3K/AKT pathway. Blocking the PI3K/AKT pathway could cancel the effect of mi R-19 a.CONCLUSION: All together, our results suggest that mi R-19 a could be used as a promising therapeutic target in the treatment of GC.展开更多
AIM:To evaluate the effect of bone morphogenetic protein-6(BMP-6)on transforming growth factor(TGF)-β_(2)-induced epithelial-mesenchymal transition(EMT)in retinal pigment epithelium(RPE).METHODS:Adult retinal pigment...AIM:To evaluate the effect of bone morphogenetic protein-6(BMP-6)on transforming growth factor(TGF)-β_(2)-induced epithelial-mesenchymal transition(EMT)in retinal pigment epithelium(RPE).METHODS:Adult retinal pigment epithelial cell line(ARPE-19)were randomly divided into control,TGF-β_(2)(5μg/L),and BMP-6 small interfering RNA(siRNA)group.The cell morphology was observed by microscopy,and the cell migration ability were detected by Transwell chamber.The EMT-related indexes and BMP-6 protein levels were detected by Western blotting.Furthermore,a BMP-6 overexpression plasmid was constructed and RPE cells were divided into the control group,TGF-β_(2)+empty plasmid group,BMP-6 overexpression group,and TGF-β_(2)+BMP-6 overexpression group.The EMT-related indexes and extracellular regulated protein kinases(ERK)protein levels were detected.RESULTS:Compared with the control group,the migration of RPE cells in the TGF-β_(2) group was significantly enhanced.TGF-β_(2) increased the protein expression levels ofα-smooth muscle actin(α-SMA),fibronectin and vimentin but significantly decreased the protein levels of E-cadherin and BMP-6(P<0.05)in RPE.Similarly,the migration of RPE cells in the BMP-6 siRNA group was also significantly enhanced.BMP-6 siRNA increased the protein expression levels ofα-SMA,fibronectin and vimentin but significantly decreased the protein expression levels of E-cadherin(P<0.05).Overexpression of BMP-6 inhibited the migration of RPE cells induced by TGF-β_(2) and prevented TGF-β_(2) from affecting EMT-related biomarkers(P<0.05).CONCLUSION:BMP-6 prevents the EMT in RPE cells induced by TGF-β_(2),which may provide a theoretical basis for the prevention and treatment of proliferative vitreoretinopathy.展开更多
文摘AIM To evaluate the clinical properties of three subpopulations of circulating tumor cells(CTCs) undergoing epithelial-mesenchymal transition(EMT) in pancreatic ductal adenocarcinoma(PDAC) patients.METHODS We identified CTCs for expression of the epithelial cell marker cytokeratin or epithelial cell adhesion molecule(EpCAM)(E-CTC), the mesenchymal cell markers vimentin and twist(M-CTC), or both(E/M-CTC) using the CanPatrol system. Between July 2014 and July 2016, 107 patients with PDAC were enrolled for CTC evaluation. CTC enumeration and classification were correlated with patient clinicopathological features and outcomes.RESULTS CTCs were detected in 78.5% of PDAC patients. The number of total CTCs ranged from 0 to 26 across all 107 patients, with a median value of six. CTC status correlated with lymph node metastasis, TNM stage, distant metastasis, blood lymphocyte counts, and neutrophil-to-lymphocyte ratio(NLR). Kaplan-Meier survival analysis showed that patients with ≥ 6 total CTCs had significantly decreased overall survival and progression-free survival compared with patients with < 6 total CTCs. The presence of M-CTCs was positively correlated with TNM stage(P < 0.01) and distant metastasis(P < 0.01). Additionally, lymphocyte counts and NLR in patients without CTCs were significantly different from those in patients testing positive for each CTC subpopulation(P < 0.01).CONCLUSION Classifying CTCs by EMT markers helps to identify the more aggressive CTC subpopulations and provides useful evidence for determining a suitable clinical approach.
文摘目的:探讨姜黄素(curcumin,Cur)抑制胰腺癌细胞上皮间质转化(epithelial-mesenchymal transition,EMT)及其影响胰腺癌侵袭转移的潜在机制.方法:常规培养3种不同分化特征的胰腺癌细胞株PANC-1、AsPC-1、BxPC-3细胞,采用Western blot法检测其EMT标志物E-cadherin、Vimentin蛋白的表达.以终浓度10 ng/mL的转化生长因子-β1(transforming growth factor beta1,TGF-β1)处理低分化PANC-1细胞24 h后,在倒置显微镜下观察其形态变化.分别以10 ng/mL TGF-β1、10μmol/L Cur+10 ng/mL TGF-β1、10μmol/L Cur处理PANC-1细胞,另设空白对照组,分别采用Real-time PCR及Western blot法检测各组E-cadherin、Vimentin表达变化.结果:Western blot结果显示,PANC-1、AsPC-1、BxPC-3细胞的E-cadherin蛋白表达依次增强(1.00±00、1.36±0.04、2.14±0.06,P<0.05),而Vimentin表达则依次减弱(1.00±0.00、0.60±0.05、0.49±0.04,P<0.05),这表明低分化的PANC-1细胞间质特性最强.TGF-β1刺激PANC-1细胞发生典型的EMT形态变化.Real-time PCR和Western b l o t结果均显示,PA N C-1细胞经姜黄素及T G F-β1处理后,与对照组相比,T G F-β1组E-cadherin mRNA及蛋白表达均明显下调(分别为0.67±0.05、0.47±0.05,P<0.05 vs对照组),Vimentin基因mRNA及蛋白则明显上调(分别为2.38±0.14、1.43±0.07,P<0.05 vs对照组),说明TGF-β1促进PANC-1细胞EMT过程.而与TGF-β1组相比,Cur+TGF-β1及Cur组的E-cadherin表达具有上调趋势(mRNA分别为0.98±0.06、1.34±0.08,P<0.05 vs TGF-β1组;蛋白为0.32±0.04、0.68±0.06,P<0.05 vs TGF-β1组),Vimentin却下调(mRNA分别为0.63±0.08、0.99±0.07,P<0.05 vs TGF-β1组;蛋白分别为1.01±0.14、0.57±0.06,P<0.05vs TGF-β1组),其蛋白与mRNA表达结果基本一致.这表明姜黄素具有阻断TGF-β1诱导的PANC-1细胞EMT效应.结论:姜黄素能够阻断TGF-β1诱导的PANC-1细胞的EMT过程,从而抑制其侵袭转移.
文摘Extracellular vesicles from highly metastatic tumor cells have been shown to mediate epithelial-mesenchymal transition(EMT)-related events in recipient cells.In endometrial epithelial cells,EMT processes are known to be involved in the development of adenomyosis.We aimed to investigate whether adenomyosis-derived extracellular vesicles(AMEVs)are able to induce an EMT process in endometrial epithelial cells.In this study,AMEVs were isolated from patients with adenomyosis and characterized by transmission electron microscopy,Western blot,and nanoparticle tracking.Primary endometrial epithelial cells(EECs)were derived from normal endometrium tissues from patients with leiomyoma and co-cultured with AMEVs in vitro.AMEV uptake was examined by fluorescence confocal microscopy.The invasion of EECs was confirmed by Transwell assay.Immunohistochemistry,Western blot,and qRT-PCR were performed on EECs to illustrate the expression levels of cytokeratin 19,E-cadherin,vimentin,and zinc finger E-boxbinding homeobox 1(ZEB1).The results indicated that the cellular fluorescence intensity gradually increased after 48 h of co-culture,but decreased after 72 h.After co-culturing with AMEVs for 72 h,EECs expressed significantly lower levels of cytokeratin 19 and E-cadherin,and significantly higher levels of vimentin and ZEB1.Together these results demonstrated that AMEVs induce an EMT process and enhance the invasion of EECs.These changes may contribute to the pathogenesis and progression of adenomyosis.
基金Supported by National Science Foundation of China,No.31471147
文摘Gastric cancer(GC) is the fifth most common malignancy in the world. The major cause of GC is chronic infection with Helicobacter pylori(H. pylori). Infection with H. pylori leads to an active inflammatory microenvironment that is maintained by immune cells such as T cells, macrophages, natural killer cells, among other cells. Immune cell dysfunction allows the initiation and accumulation of mutations in GC cells, inducing aberrant proliferation and protection from apoptosis. Meanwhile, immune cells can secrete certain signals, including cytokines, and chemokines, to alter intracellular signaling pathways in GC cells. Thus, GC cells obtain the ability to metastasize to lymph nodes by undergoing the epithelial-mesenchymal transition(EMT), whereby epithelial cells lose their epithelial attributes and acquire a mesenchymal cell phenotype. Metastasis is a leading cause of death for GC patients, and the involved mechanisms are still under investigation. In this review, we summarize the current research on how the inflammatory environment affects GC initiation and metastasis via EMT.
文摘AIM: To investigate the mechanism by which mi R-19 a is up-regulated in gastric cancer(GC), which plays an oncogenic role.METHODS: In the present study, we investigated the role of mi R-19 a in gastric tissues as well as two GC cell lines. In vivo, we detected the basal expression level of mi R-19 a using real-time reverse transcription-PCR(RTPCR), and the relevance between expression of mi R-19 a and clinicopathological information was analyzed.In vitro, mi R-19 a was ectopically expressed using overexpression and knock-down strategies.RESULTS: Overexpression of mi R-19 a was significantly associated with metastasis of GC and inferior overall prognosis. However, no significant correlation was f o u n d b e t w e e n m i R- 1 9 a e x p r e s s i o n a n d o t h e r characteristics such as age, gender, tobacco, alcohol or tumor size. Cell proliferation, migration and invasion assays showed that overexpression of mi R-19 a promoted the proliferation, migration and invasion, and that overexpression of mi R-19 a promoted the epithelialmesenchymal transition through activating the PI3K/AKT pathway. Blocking the PI3K/AKT pathway could cancel the effect of mi R-19 a.CONCLUSION: All together, our results suggest that mi R-19 a could be used as a promising therapeutic target in the treatment of GC.
基金Supported by the Key Research&Development Program of Shaanxi Province(No.2022SF-311,No.2024SFYBXM-328,No.2024SF-YBXM-325)the Natural Science Basic Research Program of Shaanxi Province,China(No.2021JQ-385).
文摘AIM:To evaluate the effect of bone morphogenetic protein-6(BMP-6)on transforming growth factor(TGF)-β_(2)-induced epithelial-mesenchymal transition(EMT)in retinal pigment epithelium(RPE).METHODS:Adult retinal pigment epithelial cell line(ARPE-19)were randomly divided into control,TGF-β_(2)(5μg/L),and BMP-6 small interfering RNA(siRNA)group.The cell morphology was observed by microscopy,and the cell migration ability were detected by Transwell chamber.The EMT-related indexes and BMP-6 protein levels were detected by Western blotting.Furthermore,a BMP-6 overexpression plasmid was constructed and RPE cells were divided into the control group,TGF-β_(2)+empty plasmid group,BMP-6 overexpression group,and TGF-β_(2)+BMP-6 overexpression group.The EMT-related indexes and extracellular regulated protein kinases(ERK)protein levels were detected.RESULTS:Compared with the control group,the migration of RPE cells in the TGF-β_(2) group was significantly enhanced.TGF-β_(2) increased the protein expression levels ofα-smooth muscle actin(α-SMA),fibronectin and vimentin but significantly decreased the protein levels of E-cadherin and BMP-6(P<0.05)in RPE.Similarly,the migration of RPE cells in the BMP-6 siRNA group was also significantly enhanced.BMP-6 siRNA increased the protein expression levels ofα-SMA,fibronectin and vimentin but significantly decreased the protein expression levels of E-cadherin(P<0.05).Overexpression of BMP-6 inhibited the migration of RPE cells induced by TGF-β_(2) and prevented TGF-β_(2) from affecting EMT-related biomarkers(P<0.05).CONCLUSION:BMP-6 prevents the EMT in RPE cells induced by TGF-β_(2),which may provide a theoretical basis for the prevention and treatment of proliferative vitreoretinopathy.
