【摘要】 目的:探讨miR-520b和抗细胞凋亡因子(DAD1)对肾癌细胞增殖、迁移和侵袭的影响及分子机制。方法:qRT-PCR和Western blot实验检测A498人肾癌细胞中miR-520b和DAD1表达。将miR-520b mimics或DAD1 siRNA转染至A498细胞,MTT和Transwell实验检测细胞活力、迁移和侵袭。Targetscan在线预测、双荧光素酶报告基因和Western blot实验验证miR-520b和DAD1的靶向关系。将miR-520b mimics和pcDNA-DAD1共转染至A498细胞,检测细胞活力、迁移和侵袭。结果:在A498细胞中,miR-520b表达下调而DAD1表达上调。在A498细胞中过表达miR-520b或敲减DAD1,细胞活力下降,迁移和侵袭细胞数减少。Targetscan在线预测、双荧光素酶报告基因和Western blot实验结果表明,miR-520b可负调控DAD1蛋白表达。过表达DAD1可逆转miR-520b对A498细胞增殖、迁移和侵袭的抑制作用。结论:miR-520b能够通过靶向调节DAD1表达抑制A498细胞增殖、迁移和侵袭。更多还原

【Abstract】 Objective:To investigate the effects and mechanism of miR-520 b and defender against cell death 1(DAD1)on proliferation,migration and invasion of renal carcinoma cells.Methods:The expression of miR-520 b and DAD1 in A498 human renal carcinoma cells were detected by qRT-PCR and Western blot,respectively.Cell viability,migration and invasion of A498 cells after transfection of miR-520 b mimics or DAD1 siRNA were detected by MTT and Transwell assays,respectively.The targeting relationship between miR-520 b and DAD1 was verified by Targetscan online prediction,dual-luciferase assay and Western blot,respectively.There was co-transfection of miR-520 b mimics and pcDNA-DAD1 into A498 cells,and cell viability,migration and invasion were detected.Results:In A498 cells,the expression of miR-520 b was down-regulated while expression of DAD1 was up-regulated.Cell viability,migration and invasion of A498 cells were decreased after overexpression of miR-520 b or knockdown of DAD1.The results of Targetscan online prediction,dual-luciferase assay and Western blot indicted that miR-520 b could negatively regulate the protein expression of DAD1.Overexpression of DAD1 could reverse the inhibition of miR-520 b on proliferation,migration and invasion of A498 cells.Conclusion:miR-520 b could inhibit proliferation,migration and invasion of A498 cells by targeting expression of DAD1.更多还原