miR-17-5p靶向PTEN蛋白调节EMT抑制胃癌细胞的增殖

miR-17-5p targeting PTEN protein regulates EMT to inhibit the proliferation of gastric cancer cells

目的 检测miR-17-5p在胃癌组织及胃癌细胞系中的表达及其对N-cadherin和Vimentin表达的影响。方法 RT-PCR方法检测胃癌组织及胃癌细胞中miR-17-5p的表达情况;将miR-17-5p inhibitor转染至胃癌细胞,RT-PCR方法验证转染效率,CCK8方法检测抑制miR-17-5p对胃癌细胞增殖能力的影响,transwell及划痕实验探究抑制miR-17-5p对胃癌细胞侵袭和迁移能力的影响。建立移植瘤模型,观察抑制miR-17-5p对胃癌细胞体内肿瘤生长的影响,免疫荧光实验检测移植瘤EMT相关蛋白N-cadherin和Vimentin的表达情况。生物信息学方法预测miR-17-5p与PTEN的潜在结合位点,荧光素酶报告基因实验验证PTEN是miR-17-5p的下游靶蛋白。结果 与癌旁组织及上皮细胞相比,miR-17-5p在胃癌组织和胃癌细胞中呈现高表达。抑制miR-17-5p可降低胃癌细胞增殖、迁移和侵袭能力、降低胃癌细胞移植瘤的体积和质量、下调N-cadherin和Vimentin蛋白表达。生物信息学预测PTEN是miR-17-5p的下游靶蛋白,荧光素酶报告基因实验也证实了这一结果。结论 miR-17-5p在胃癌组织及胃癌细胞中呈现高表达,与EMT相关N-cadherin和Vimentin蛋白表达呈正相关,可能参与促进胃癌发展。

Objective To study the expression of miR-17-5p in gastric cancer tissues and gastric cancer cell lines and its effect on the expression of N-cadherin and Vimentin.Methods The expression of miR-17-5p in gastric cancer cells was determined by RT-PCR.miR-17-5p inhibitor was transfected into gastric cancer cells to establish the transfection efficiency was verified by RT-PCR,and the effect of miR-17-5p was determined by CCK8.The effect of the inhibition of miR-17-5p on the invasion and migration ability of gastric cancer cells was explored by transwell and scratch experiments.The effect of miR-17-5p inhibition on the growth of gastric cancer cells in vivo was observed,and the expression of EMT-related proteins N-cadherin and Vimentin in the transplanted tumor was detected by immunofluorescence assay.Bioinformatics methods were used to predict the potential binding sites of miR-17-5p and PTEN,and luciferase reporter gene experiments verified that PTEN was the downstream target protein of miR-17-5p.Results miR-17-5p showed highly expression in gastric cancer tissues and epithelial cells.Inhibition of miR-17-5p decreased the value-added activity,migration and invasion ability of gastric cancer cells,it can also reduce the volume,mass,the expression of N-cadherin and Vimentin index proteinof gastric cancer cells.Bioinformatics predicted that PTEN was a downstream target of miR-17-5p,and these results were also confirmed by the luciferase reporter gene assay.Conclusion miR-17-5p is highly expressed in gastric cancer tissues and cells,and is positively correlated with EMT related N-cadherin and Vimentin protein expression,which may participate in promoting the development of gastric cancer.