miR－940抑制Cbl－b的表达促进胃癌细胞的增殖

摘要

Objective:To investigate the effect of the proliferation of miR－940 on gastric cancer cell lines,and to explore the role of miR－940 and Cbl－b signal pathways in this process.Methods:Real－time PCR detected the ex-pression of miR－940 on gastric cancer cell lines.MTT detected the proliferation of gastric cancer cells.Dual lucifer-ase reporter assay detected the bingding ability between miR－940 and Cbl－b.The expression of the protein was tes-tified by Western blotting analysis.Results:MTT showed that miR－940 promoted the proliferation of MGC803.A pu-tative seed sequence of miR－940 was found in the 3'UTR of the Cbl－b transcript.Dual luciferase reporter assay showed that Cbl－b was the target gene of miR－940.Western blotting showed that mimic miR－940 inhibited the ex-pression of Cbl－b.Cbl－b suppressed the proliferation of MGC803.miR－940 promoted the proliferation of MGC803 through inhibiting the expression of Cbl－b.Conclusion:miR－940 inhibited the proliferation of MGC803 through in-hibiting the expression of Cbl－b.%目的:研究miR－940对胃癌细胞增殖的影响,探讨miR－940和Cbl－b信号转导通路在此过程中的作用.方法:采用Real－time PCR法检测胃癌细胞中miR－940的表达,MTT法检测胃癌细胞的增殖能力,双荧光素酶报告基因检测系统检测miR－940与Cbl－b的结合,Western blotting实验观察蛋白的表达.结果:MTT法显示miR－940可促进胃癌细胞MGC803的增殖,BLAST对比分析结果显示Cbl－b与miR－940存在结合位点.双荧光素酶报告基因检测系统证实Cbl－b是miR－940的靶基因.Western blotting结果显示过表达miR－940后,Cbl－b的表达明显下调.Cbl－b抑制胃癌细胞MGC803的增殖.miR－940通过负向调节Cbl－b的表达促进胃癌细胞的增殖.结论:miR－940通过抑制Cbl－b的表达,促进胃癌细胞MGC803的增殖.