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目的探讨中介体复合物亚基19(Med19)对人膀胱癌UM-UC3细胞迁移能力的影响及其机制。方法采用针对Med19的siRNA慢病毒载体转染人膀胱癌UM-UC3,应用实时荧光定量PCR(qRT-PCR)和蛋白质印迹方法(Western blot)检测Med19-siRNA转染组(siRNA)与空载组(NC)Medl9基因表达情况;采用Western blot检测细胞外基质金属蛋白酶(MMP)-2和MMP-9的表达;采用划痕实验和Transwell实验检测细胞迁移能力的变化。结果 Medl9-siRNA慢病毒感染UM-UC3细胞后,转染组Medl9mRNA表达与空载组相比明显降低,差异有统计学意义(t=10.15,P<0.01),且转染组Med19、MMP-2和MMP-9蛋白表达明显降低,差异有统计学意义(t值分别为71.36,26.34,8.627,均P<0.01),划痕实验和Transwell实验显示转染组细胞迁移能力明显减弱。结论沉默Med19基因可通过抑制MMP-2和MMP-9的表达降低人膀胱癌细胞UM-UC3的迁移能力。
Objective To investigate the effect of Med19 on the migration of human bladder cancer UM-UC3 cells and its mechanism. Methods Human bladder cancer cell line UM-UC3 was transfected with siRNA targeting Med19. Real-time quantitative PCR (qRT-PCR) and Western blot were used to detect the expression of Med19-siRNA transfection group (siRNA) (NC) Medl9 gene expression; Western blot detection of extracellular matrix metalloproteinase (MMP) -2 and MMP-9 expression; using scratch test and Transwell assay of cell migration ability changes. Results After Medl9-siRNA lentivirus infected UM-UC3 cells, the expression of Medl9 mRNA in transfected group was significantly lower than that in empty vector group (t = 10.15, P <0.01) 2 and MMP-9 protein expression was significantly lower (t = 71.36,26.34,8.627, respectively, P <0.01). Scratch assay and Transwell assay showed that the transfected group of cells migrated significantly weakened. Conclusion Silencing Med19 can reduce the migration of human bladder cancer cell line UM-UC3 by inhibiting the expression of MMP-2 and MMP-9.