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目的:探讨hsa-miR-99b在胃腺癌细胞系的表达及其影响胃腺癌细胞系增殖和迁移的潜在作用机制。方法:Real-time PCR检测miR-99b在人胃上皮细胞系GES-1和人胃腺癌细胞系SGC-7901、BGC-823、MGC-803中的表达。用人工合成的miR-99b模拟物转染SGC-7901细胞构建过表达miR-996细胞系,转染对照模拟物作为对照组,CCK-8法检测过表达miR-99b的SGC-7901细胞的增殖情况,Transwell法检测SGC-7901细胞系的迁移情况,Western blotting检测SGC-7901细胞系中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)的表达情况。结果:miR-99b在胃腺癌细胞系中的表达水平显著低于胃上皮细胞系,其中以SCG-7901细胞最甚。转染miR-99b模拟物48 h后,实验组SCG-7901细胞增殖能力低于对照组(0.20±0.00 vs 0.27±0.02,P<0.05);与转染对照模拟物的细胞相比,转染24 h后,实验组SCG-7901穿膜细胞数低于对照组[(226.67±25.17)vs(523.33±25.17)个,P<0.05]。miR-99b可下调人胃腺癌细胞系SGC-7901中m TOR蛋白的表达。结论:miR-99b在胃腺癌细胞系中的表达明显下调,miR-99b抑制人胃腺癌细胞系SGC-7901增殖和迁移的作用可能与下调m TOR表达有关。
Objective: To investigate the expression of hsa-miR-99b in gastric adenocarcinoma cell lines and its potential mechanism of action in the proliferation and migration of gastric adenocarcinoma cell lines. Methods: Real-time PCR was used to detect the expression of miR-99b in human gastric epithelial cell line GES-1 and human gastric adenocarcinoma cell lines SGC-7901, BGC-823 and MGC-803. The synthetic miR-99b mimics were transfected into SGC-7901 cells to construct overexpression miR-996 cell line, and the control mimics were transfected as control group. The proliferation of SGC-7901 cells overexpressing miR-99b was detected by CCK-8 assay Transwell method was used to detect the migration of SGC-7901 cell line. The expression of mammalian target of rapamycin (mTOR) in SGC-7901 cell line was detected by Western blotting. Results: The expression level of miR-99b in gastric adenocarcinoma cell lines was significantly lower than that of gastric epithelial cell lines, of which SCG-7901 cells were the most. Compared with the control group, the proliferation of SCG-7901 cells in the experimental group was significantly lower than that in the control group (0.20 ± 0.00 vs 0.27 ± 0.02, P <0.05) after transfected with the miR-99b mimics for 48 h. Compared with the cells transfected with the control mimics, After 24 h, the number of SCG-7901 cells in the experimental group was significantly lower than that in the control group [(226.67 ± 25.17) vs (523.33 ± 25.17) P <0.05]. miR-99b down-regulates the expression of mTOR protein in human gastric adenocarcinoma cell line SGC-7901. Conclusion: The expression of miR-99b in gastric adenocarcinoma cell line was significantly down-regulated. The effect of miR-99b on proliferation and migration of human gastric adenocarcinoma cell line SGC-7901 may be related to down-regulation of mTOR expression.