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目的:探讨微小RNA145(miRNA145)对肝癌细胞生长、凋亡及侵袭转移的影响。方法:实验将Hep G2细胞随机设置为3组:空白对照组、空质粒组及转染组,转染后real-time PCR法检测验证各组细胞miRNA145及N-钙黏蛋白(N-cadherin)mRNA的表达,并用Western blot检测miRNA145目标蛋白N-钙黏蛋白的表达,MTS增殖实验检测各组细胞的生长情况,流式细胞术检测细胞周期及细胞凋亡率,Transwell法检测各组细胞的侵袭转移能力。结果:Real-time PCR结果证实转染组细胞miRNA145表达比空质粒组和空白对照组细胞明显增多(P<0.05),同时N-钙黏蛋白的表达在转染组细胞中显著减少(P<0.05);经Western blot结果证实,转染组细胞中N-钙黏蛋白表达比空质粒组和空白对照组细胞明显减少(P<0.05);转染组细胞活性在转染后第2天、第3天均明显下降,70.04%的细胞周期停止在G1期,转染后Hep G2细胞凋亡率明显增高,细胞的侵袭转移能力下降,差异均有统计学意义。结论:上调miRNA145表达能有效抑制肝癌细胞的细胞周期进展和侵袭转移,促进细胞凋亡。
Objective: To investigate the effect of microRNA 145 (miRNA 145) on the growth, apoptosis, invasion and metastasis of hepatocellular carcinoma cells. Methods: Hep G2 cells were randomly divided into three groups: blank control group, empty plasmid group and transfection group. After transfection, real-time PCR assay confirmed the expression of miRNA145 and N-cadherin The expression of miRNA145 target protein N-cadherin was detected by Western blot. The growth of each group of cells was detected by MTS proliferation assay. The cell cycle and apoptosis rate were detected by flow cytometry. Invasion and metastasis ability. Results: The results of Real-time PCR confirmed that the expression of miRNA145 in transfected cells was significantly increased (P <0.05) and the expression of N-cadherin was significantly decreased in transfected cells compared with those in empty plasmid group and blank control group (P < 0.05). The results of Western blot showed that the expression of N-cadherin in transfected cells was significantly decreased compared with that in empty plasmid group and blank control group (P <0.05) On the third day, the cell cycle was significantly decreased. The cell cycle arrest of 70.04% was in G1 phase. The apoptosis rate of Hep G2 cells was significantly increased after transfection, and the invasion and metastasis of cells decreased. The difference was statistically significant. Conclusion: Up-regulation of miRNA145 can effectively inhibit cell cycle progression, invasion and metastasis of hepatocellular carcinoma cells and promote apoptosis.