TGF-β1对食管癌上皮间质转化的调控研究
发布时间：2018-08-12 17:37
【摘要】：目的:探讨转化生长因子(transforming growth factor-β,TGF-β1)在食管癌中的作用及其对上皮间质转化(Epithelial-Mesenchymal Transition,EMT)的调控。方法:利用TGF-β1受体抑制剂LY2109761处理食管癌Eca109细胞,根据浓度梯度(0、0.1、1、10μm/ml)分为四组;慢病毒转染TGF-β1干扰RNA(siRNA)处理食管癌Eca109细胞,分为3组:实验组(病毒稳定TGF-β1 si RNA转染Eca109细胞)、阴性对照组(转染无关序列Eca109细胞)、空白对照组(正常未经任何处理的Eca109细胞),采用荧光定量PCR(Quantitative Realtime-PCR,qRT-PCR)技术检测TGF-β1及上皮间质转化(EMT)相关指标E-钙粘蛋白(E-cadherin)、波形蛋白(Vimentin)、锌指转录因子(Snail)在mRNA水平表达情况;Western blot技术检测E-cadherin、Vimentin蛋白水平的表达;同时通过细胞增殖实验(MTT)、细胞划痕实验及Transwell法分别检测细胞增殖、迁移、侵袭的变化情况。结果:通过TGF-β1受体抑制剂(0、0.1、1、10μm/ml)处理Eca109细胞后,在mRNA水平TGF-β1表达逐渐降低,E-cadherin表达逐渐升高,Vimentin、Snail表达逐渐降低(P0.05);在蛋白水平,E-cadherin表达逐渐升高,Vimentin表达逐渐降低(P0.05);此外,细胞增殖和迁移能力逐渐降低(P0.05)。通过慢病毒转染TGF-β1 si RNA后发现,实验组与空白对照组和阴性对照组相比,TGF-β1 mRNA表达降低(P0.05),E-cadherin表达增高,Vimentin表达降低;同时实验组细胞增殖、迁移和侵袭能力均降低(P0.05)。结论:TGF-β1可能通过上皮间质转化促进食管癌细胞的增殖、迁移和侵袭。
[Abstract]:Aim: to investigate the role of transforming growth factor (transforming growth factor- 尾 1 (TGF- 尾 1) in esophageal carcinoma and its regulation on Epithelial-Mesenchymal transition. Methods: esophageal carcinoma Eca109 cells were treated with TGF- 尾 1 receptor inhibitor (LY2109761) and divided into four groups according to the concentration gradient (0 0. 1 渭 m/ml), lentivirus transfected with TGF- 尾 1 interfered with RNA (siRNA) to treat esophageal cancer Eca109 cells. TGF- 尾 1 was divided into three groups: experimental group (viral stable TGF- 尾 1si RNA transfected Eca109 cells), negative control group (transfection of unrelated Eca109 cells) and blank control group (normal untreated Eca109 cells). TGF- 尾 1 was detected by fluorescence quantitative PCR (Quantitative real time PCR qRT-PCR. Ecadherin (E-cadherin) and (Vimentin), zinc finger transcription factor (Snail) (Snail) were detected by Western blot. At the same time, the cell proliferation, migration and invasion were detected by (MTT), cell scratch assay and Transwell assay. Results: after treated with TGF- 尾 1 receptor inhibitor (0 0. 1 m/ml), the expression of TGF- 尾 1 decreased gradually at the mRNA level and decreased gradually at the mRNA level (P0.05), while at the protein level, the expression of E-cadherin decreased gradually (P0.05), and the expression of Vimentin decreased gradually at the protein level (P0.05). The ability of cell proliferation and migration decreased gradually (P0.05). After lentivirus transfection of TGF- 尾 1si RNA, it was found that the expression of TGF- 尾 1 mRNA in the experimental group was lower than that in the blank control group and the negative control group (P0.05), while the expression of Vimentin in the experimental group was lower than that in the control group (P0.05), and the ability of cell proliferation, migration and invasion in the experimental group was decreased (P0.05). Conclusion TGF- 尾 1 may promote the proliferation, migration and invasion of esophageal carcinoma cells through epithelial interstitial transformation.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.1
[Abstract]:Aim: to investigate the role of transforming growth factor (transforming growth factor- 尾 1 (TGF- 尾 1) in esophageal carcinoma and its regulation on Epithelial-Mesenchymal transition. Methods: esophageal carcinoma Eca109 cells were treated with TGF- 尾 1 receptor inhibitor (LY2109761) and divided into four groups according to the concentration gradient (0 0. 1 渭 m/ml), lentivirus transfected with TGF- 尾 1 interfered with RNA (siRNA) to treat esophageal cancer Eca109 cells. TGF- 尾 1 was divided into three groups: experimental group (viral stable TGF- 尾 1si RNA transfected Eca109 cells), negative control group (transfection of unrelated Eca109 cells) and blank control group (normal untreated Eca109 cells). TGF- 尾 1 was detected by fluorescence quantitative PCR (Quantitative real time PCR qRT-PCR. Ecadherin (E-cadherin) and (Vimentin), zinc finger transcription factor (Snail) (Snail) were detected by Western blot. At the same time, the cell proliferation, migration and invasion were detected by (MTT), cell scratch assay and Transwell assay. Results: after treated with TGF- 尾 1 receptor inhibitor (0 0. 1 m/ml), the expression of TGF- 尾 1 decreased gradually at the mRNA level and decreased gradually at the mRNA level (P0.05), while at the protein level, the expression of E-cadherin decreased gradually (P0.05), and the expression of Vimentin decreased gradually at the protein level (P0.05). The ability of cell proliferation and migration decreased gradually (P0.05). After lentivirus transfection of TGF- 尾 1si RNA, it was found that the expression of TGF- 尾 1 mRNA in the experimental group was lower than that in the blank control group and the negative control group (P0.05), while the expression of Vimentin in the experimental group was lower than that in the control group (P0.05), and the ability of cell proliferation, migration and invasion in the experimental group was decreased (P0.05). Conclusion TGF- 尾 1 may promote the proliferation, migration and invasion of esophageal carcinoma cells through epithelial interstitial transformation.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.1
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