miR-1266靶向DAB2IP调控肝癌细胞的增殖和凋亡

miR-1266 targets DAB2IP to regulate the proliferation and apoptosis of hepatocellular carcinoma cells

目的探讨miR-1266对肝癌细胞增殖和凋亡的影响及其调控机制。方法选取西安市第八医院2018年1月至2018年10月收治的40例肝癌患者,利用实时荧光定量PCR(qRT-PCR)检测肝癌组织中miR-1266和DAB2IP的表达水平并分析其相关性;利用生物信息学网站预测miR-1266潜在靶基因DAB2IP,双荧光素酶实验进行验证;利用脂质体转染技术向在肝癌HepG2细胞中转染miR-1266 mimics、miR-1266 mimics+pcDNA-DAB2IP,qRT-PCR验证转染效率;利用Western blotting检测转染后细胞中DAB2IP蛋白表达水平;CCK-8实验和流式细胞仪分别检测转染后细胞的增殖和凋亡情况。结果qRT-PCR结果显示,肝癌组织中miR-1266表达上调,而DAB2IP表达下调,两者表达水平呈负相关。生物信息学预测miR-1266的靶基因可能是DAB2IP,双荧光素酶报告实验显示,DAB2IP是miR-1266的靶基因。Western blotting实验结果显示,转染miR-1266 mimics显著降低了DAB2IP蛋白表达,miR-1266 mimics+pcDNA-DAB2IP共转染恢复了DAB2IP蛋白表达。CCK-8增殖实验和流式细胞仪结果显示,转染miR-1266 mimics显著促进了HepG2细胞增殖,抑制了细胞凋亡,miR-1266 mimics+pcDNA-DAB2IP共转染逆转了上述现象。结论肝癌组织中miR-1266表达上调,DAB2IP表达下调;miR-1266通过靶向下调DAB2IP表达促进肝癌细胞增殖、抑制细胞凋亡。

Objective To investigate the effect of miR-1266 on proliferation and apoptosis of hepatocellular carcinoma cells and its regulatory mechanism.Methods Forty hepatocellular carcinoma patients were selected from Jun.2018 to Oct.2018 in the Eighth Hospital of Xi’an.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression levels of miR-1266 and DAB2 IP in hepatocellular carcinoma tissues and analyze its correlation.Bioinformatics website was used to predict the potential target gene DAB2 IP of miR-1266,and double luciferase experiment was used to verify it.Liposome transfection technology was used to transfect miR-1266 mimics,miR-1266 mimics+pcDNA-DAB2 IP into HepG2 cells,and qRT-PCR was used to verify the transfection efficiency.The expression of DAB2 IP protein was detected by Western blotting,and the proliferation and apoptosis of the transfected cells were detected by CCK-8 assay and flow cytometry respectively.Results The results of qRT-PCR showed that the expression of miR-1266 was up-regulated,while the expression of DAB2 IP was down-regulated in hepatocellular carcinoma tissues,and the expression levels of both were negatively correlated.Bioinformatics predicted that the target gene of miR-1266 might be DAB2 IP,and double luciferase report experiments showed that DAB2 IP was a target gene of miR-1266.The results of Western blotting showed that transfection of miR-1266 mimics significantly reduced DAB2 IP protein expression,and co-transfection of miR-1266 mimics+pcDNA-DAB2 IP restored DAB2 IP protein expression.The results of CCK-8 proliferation experiments and flow cytometry showed that transfection of miR-1266 mimics significantly promoted HepG2 cell proliferation and inhibited apoptosis,co-transfection of miR-1266 mimics+pcDNA-DAB2 IP reversed the above phenomenon.Conclusion The expression of miR-1266 is up-regulated,while the expression of DAB2 IP is down-regulated in hepatocellular carcinoma tissues;miR-1266 promotes the proliferation and inhibits apoptosis of hepatocellular carcinomacells by targeting down-regulation of DAB2 IP expression.