MicroRNA-133b抑制人横纹肌肉瘤细胞增殖与迁移的研究  

The Mechanism of MicroRNA-133b-mediated Inhibition of Human Rhabdomyosarcoma Cell Proliferation and Migration

作　　者：王丽花[1] 陈通克[2] 姚莎莎[1] 陈晓燕[1] 王教[1] 

机构地区：[1]温州医学院眼视光学院,温州325027 [2]温州医学院实验动物中心,温州325000

出　　处：《中国细胞生物学学报》2012年第5期461-467,共7页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金(No.Y2080853);温州医学院眼视光医院创新引导课题(No.YNCX201102)资助项目~~

摘　　要：该研究首先通过Real-time RT-PCR检测发现,microRNA-133b在人横纹肌肉瘤细胞RD和A204中的表达量比正常肌肉组织中的表达量明显降低,再通过阳离子脂质体介导的方法将microRNA-133b转染入横纹肌肉瘤细胞RD和A204细胞中,并应用MTS法、Transwell法研究发现,microRNA-133b可明显抑制RD和A204细胞的增殖与迁移;采用Western blot法检测转染后细胞内与增殖、迁移及细胞周期相关的蛋白表达,发现microRNA-133b能下调RD和A204细胞中的LIMand SH3 protein 1(LASP1)、c-MET、p-MET、p-AKT、p-ERK1/2、p-Rb的表达水平,并降低RD和A204细胞中细胞周期蛋白CDK4和CDK6表达量。该研究表明,microRNA-133b是通过下调LASP1、c-MET和p-MET的表达水平,影响c-MET下游信号分子p-AKT、p-ERK1/2的表达水平,同时还下调细胞周期相关蛋白如CDK4、CDK6、p-Rb等的表达,从而影响RD和A204细胞的增殖与迁移。The Real-time RT-PCR assay showed microRNA-133b was highly expressed in human muscle tissues but downregulated in rhabdomyosarcoma cell line RD and A204. MicroRNA-133b was transfected into RD and A204 cells by lipofectamine. The proliferation of RD and A204 cells was examined by MTS cell proliferation assay and the migration of RD and A204 cells was examined by Transwell migration assay. The results showed that microRNA-133b can inhibit the proliferation and migration of RD and A204 cells. The expression level of cell proliferation, migration and cell cycle related proteins was determined by Western blot. MicroRNA-133b down- regulated the expression of LASP1, c-MET, phosphorylated-MET, phosphorylated-AKT, phosphorylated-ERK1/2, phosphorylated-Rb, CDK4 and CDK6 in RD and A204 cells. These results indicate that microRNA-133b inhibits proliferation and migration of rhabdomyosarcoma cells through down-regulation of LASP1, c-MET, phosphorylated-MET, phosphorylated-AKT, phosphorylated-ERK1/2, phosphorylated-Rb, CDK4 and CDK6.

关 键 词：MicroRNA-133b 横纹肌肉瘤 增殖 迁移 细胞周期 

分 类 号：R738.7[医药卫生—肿瘤]