摘要
该文主要探讨Mirtron类micro RNA6894-5p过表达对胃癌细胞迁移、侵袭和增殖的影响。将miRNA6894-5p的模拟物分别转染进入胃癌MGC803和SGC7901细胞中,构建miRNA6894-5p过表达的细胞系;实时荧光定量PCR测miRNA6894-5p的RNA表达;Transwell实验检测细胞迁移侵袭能力;Cell Counting Kit 8实验检测细胞的增殖能力;荧光素酶报告基因实验检测miRNA6894-5p与肝配蛋白A3的靶向关系;实时荧光定量PCR和蛋白质印迹法进一步检测miRNA6894-5p过表达后EFNA3的m RNA和蛋白的变化。结果显示,成功构建miRNA6894-5p过表达模型的胃癌细胞系;与相应阴性对照组相比,过表达miRNA6894-5p可提高细胞迁移侵袭能力(P<0.05)和增强细胞增殖能力(P<0.05);荧光素酶报告基因实验证实,miRNA6894-5p靶向作用于肝配蛋白A3,过表达miRNA6894-5p后肝配蛋白A3的m RNA及蛋白水平显著下降(P<0.05)。该研究结果显示,Mirtron类miRNA6894-5p在人胃癌细胞中过表达可促进胃癌细胞的迁移、侵袭和增殖。
This article was to investigate the effects of human micro RNA6894-5 p(miRNA6894-5 p) on migration, invasion and proliferation of human gastric carcinoma cells. Mimics of miRNA6894-5 p were constructed and transfected into gastric cancer cells. The expression of miRNA6894-5 p in gastric carcinoma cells after transfection was determined by quantitative real time PCR(q RT-PCR). The migration, invasion and proliferation ability of gastric cancer cells were detected by Transwell chamber test and Cell Counting Kit-8 assays respectively. Luciferase reporter gene assay was used to detect the targeted relationship of miRNA6894-5 p and hepatic protein A3. The m RNA and protein expression of hepatic protein A3 were detected by q RT-PCR and Western blot. The results showed that micro RNA6894-5 p mimics were successfully transfected into gastric carcinoma cells. Compared with control cells, the ability of migration and invasion were significantly increased in overexpression cells(P<0.05) and miRNA6894-5 p overexpression could significantly promote gastric cancercells proliferation(P<0.05). The relationship between miRNA6894-5 p and EFNA3 was confirmed by Luciferase reporter gene assay. The m RNA and protein expression of EFNA3 were downregulated in miRNA6894-5 p overexpression cell. The above results indicate that miRNA6894-5 p can promote the migration, invasion and proliferation of gastric cancer cells.
引文
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