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Received:April 07, 2019 Published Online:September 20, 2019
Received:April 07, 2019 Published Online:September 20, 2019
中文摘要: 目的探讨脱氧胆酸(DCA)与G蛋白耦联胆汁酸受体5(TGR-5)在胆管癌细胞增殖及转移发生过程中的作用。方法通过PCR及Western blot检测TGR-5在胆管癌细胞RBE及正常胆管细胞HIBEPic中的表达水平;采用MTT检测不同浓度DCA对RBE细胞增殖的影响;采用Transwell检测DCA对RBE细胞侵袭力的影响;采用Western blot检测DCA对RBE细胞中上皮细胞间质化(EMT)相关蛋白[上皮细胞标志物:上皮钙黏蛋白(E-cadherin),间叶组织标志物:神经钙黏蛋白(N-cadherin)和波形蛋白(Vimentin),转录因子Snail]的表达水平。结果TGR-5在胆管癌细胞RBE中高表达,在正常胆管细胞HIBEPic中低表达;DCA能够提高胆管癌细胞的活力,且随DCA浓度的递增而增升(P<0.01);Transwell结果显示与未加DCA的RBE细胞相比,加DCA的RBE细胞中侵袭细胞数目明显增多;Western blot实验结果提示加DCA的RBE细胞的E-cadherin表达下调,而N-cadherin、Vimentin和Snail表达上调。结论TGR-5在胆管癌RBE细胞中高表达,可能与胆管癌的进展密切相关。DCA能够增强RBE细胞活力,促进其增殖。DCA能通过上调Snail、N-cadherin、Vimentin表达,下调E-cadherin表达,可能通过诱导RBE细胞发生EMT,促进胆管癌细胞转移。
中文关键词: 胆管癌 脱氧胆酸 G蛋白耦联胆汁酸受体5 上皮细胞间质化
Abstract:Objective To investigate the impacts of deoxycholic acid (DCA) and Takeda-G protein-coupled bile acid receptor 5 (TGR-5) on the proliferation and metastasis of cholangiocarcinoma cells. Methods The expressions of TGR-5 in human cholangiocarcinoma cells (RBE) and normal intrahepatic biliary epithelial cells (HIBEPic) were detected by polymerase chain reaction(PCR) and Western blot.Using MTT method,the effect of different concentrations of DCA on the proliferation of RBE cells was measured.Transwell was used to detect the effect of DCA on the invasiveness of RBE cells.Western blot was used to determine the expression levels of epithelial mesenchymal transition(EMT)-related proteins(epithelial cell markers:E-cadherin, mesenchymal tissue markers: N-cadherin ,Vimentin and transcription factor Snail)in RBE cells. Results TGR-5 was highly expressed in cholangiocarcinoma RBE cells and lowly expressed in HIBEPic lines.DCA could increased the viability of cholangiocarcinoma cells,and increased with the increase of DCA concentration.Transwell results showed that the number of invasive cells in RBE cells with 400 μM DCA was significantly higher than that in RBE cells without DCA.Western blot results suggested that down-regulated expression of E-cadherin and up-regulated expressions of N-cadherin,Vimentin and Snail induced EMT in RBE cells without DCA. Conclusion The high expression of TGR-5 in RBE cells of cholangiocarcinoma is closely related to the progression of cholangiocarcinoma.DCA can enhance RBE cell viability and promote its proliferation.By up-regulating the expressions of Snail,N-cadherin and Vimentin and down-regulating the expression of E-cadherin,DCA can induce EMT in RBE cells and promote the metastasis of cholangiocarcinoma cells.
keywords: Cholangiocarcinoma Deoxycholic acid Takeda-G protein-coupled bile acid receptor 5 Epithelial mesenchymal transition
文章编号: 中图分类号: 文献标志码:A
基金项目:南京市医学科技发展计划(YKK14176)
Author Name | Affiliation |
CHEN Shuang,ZHENG Shi-qin,WANG Xiao-song,ZHANG Xiu-hua | Center of Digestive Medicine,Second Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210011,China |
Author Name | Affiliation |
CHEN Shuang,ZHENG Shi-qin,WANG Xiao-song,ZHANG Xiu-hua | Center of Digestive Medicine,Second Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210011,China |
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