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目的研究micro RNA-126(mi R-126)对肝癌细胞SMMC-7721迁移能力的影响以及对靶基因PIK3R2表达的影响。方法通过慢病毒转染肝癌细胞株SMMC-7721使其过表达mi R-126,利用CCK-8法检测细胞增殖能力。流式细胞术检测细胞凋亡;采用实时定量PCR和蛋白印迹检测细胞PIK3R2表达水平的变化,并通过荧光素酶实验验证mi R-126与PIK3R2基因的直接调控关系。结果与对照组相比,转染mi R-126组的SMMC-7721细胞的增殖能力减弱,凋亡增加。过表达mi R-126后,SMMC-7721细胞PIK3R2的蛋白表达下调(P=0.0135)。荧光素酶报告基因实验显示,mi R-126能明显抑制PIK3R2-3’UTR的荧光素酶活性(P=0.0016)。结论过表达mi R-126可能通过靶向降低PIK3R2基因的蛋白表达,抑制肝癌细胞的迁移。
Objective To investigate the effect of micro RNA-126 (mi R-126) on the migration of hepatocellular carcinoma cell line SMMC-7721 and its effect on the expression of PIK3R2 gene. Methods The lentivirus was transfected into hepatoma cell line SMMC-7721 to overexpress mi R-126. Cell proliferation was determined by CCK-8 assay. Flow cytometry was used to detect apoptosis. The changes of PIK3R2 expression were detected by real-time quantitative PCR and Western blotting. The direct regulation of miR-126 and PIK3R2 gene was verified by luciferase assay. Results Compared with the control group, the proliferation of SMMC-7721 cells transfected with mi R-126 group decreased and the apoptosis increased. After overexpression of mi R-126, the protein expression of PIK3R2 in SMMC-7721 cells was down-regulated (P = 0.0135). Luciferase reporter assay showed that mi R-126 significantly inhibited the luciferase activity of PIK3R2-3’UTR (P = 0.0016). Conclusion Overexpression of mi R-126 may decrease the protein expression of PIK3R2 gene and inhibit the migration of hepatoma cells.