沉默中介体复合物亚基19基因对人膀胱癌细胞UM-UC3迁移能力的影响

Lentivirus-mediated inhibition of Med19 suppresses migration of bladder cancer UM-UC3 cells

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 study the effect of lentivirus-mediated inhibition of Med19 on the migration of human bladder cancer UM-UC3 cells,and to explore its potential mechanism.Methods The lentivirus vectors containing small interfering RNA（siRNA）targeting Medl9 gene were constructed and transfected to human bladder cancer UM-UC3 cells.Quantitative real-time PCR（qRT-PCR）and Western blot analysis were used to detect the Medl9 expression in the siRNA group（LvMed19siRNA cells）and the NC group（Lv-NC-infected cells）72 h after the transfection.Western blot was used to examine the changes of expression in matrix metalloproteinase 2（MMP-2）and MMP-9.The influence of Medl9 on the migration of bladder cancer cell was assessed with wound healing assay and transwell assay.Results The expressions of MMP-2 and MMP-9 were greatly reduced72 h after Med19inhibition（t value was 26.34 and8.627,P〈0.01）,and the migration ability was remarkably inhibited in the siRNA group.Conclusion Med19 inhibition reduces migration ability of human bladder cancer UM-UC3 cells via down-regulating MMP-2 and MMP-9 expressions.