RNA干扰c-maf基因表达对多发性骨髓瘤细胞凋亡的影响及机制研究

Effect and Mechanism of RNA Interference of c-Maf on the Apoptosis of Multiple Myeloma Cells

目的:探讨RNA干扰c-maf基因表达对多发性骨髓瘤细胞凋亡的影响及机制。方法:将人骨髓瘤细胞株NCI-H929分为正常对照组、转染si RNA Control的阴性对照组和转染si RNA c-maf基因的沉默组,转染48 h后,提取细胞中的蛋白,Western blot检测各组细胞中c-maf的蛋白表达,流式细胞仪检测细胞凋亡情况,Western blot检测凋亡相关蛋白Cleaved caspase3及Wnt/β-catenin信号通路相关蛋白β-catenin和Cyclin D1蛋白表达。结果:阴性对照组c-maf的蛋白表达与正常对照组比较差异无统计学意义(P>0.05),沉默组c-maf的蛋白表达显著低于正常对照组(P<0.01),而阴性对照组c-maf的蛋白表达与正常对照组比较差异无统计学意义(P>0.05)。与阴性对照组比较,沉默组的细胞凋亡率显著升高,Cleaved caspase3蛋白显著上调表达,β-catenin和Cyclin D1蛋白显著下调表达(P<0.01)。结论:RNA干扰c-maf基因表达可诱导多发性骨髓瘤细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关。

Objective： To investigate the effect and mechanism of RNA interference of c-Mar gene on the apoptosis of multiple myeloma cells. Methods： Human myeloma cell line NCI-H929 were divided into the normal control group, negative control group of transfeeted siRNA Control and silent group of siRNA c-Maf gene; cells protein were extracted after transfection for 48 h, the apoptosis was detected by flow cytometry; the expression of c-Maf, cleaved caspase3, β-catenin, Cyclin D1 protein were detected by Western blot. Results： The expression of c-Mafprotein in normal and negative control group showed no significant difference （P〉0.05）, the expression of c-Mafprotem in silent group was significantly lower than that of the normal control group （P〈0.01）; however, there was no significant difference in the expression of c-Mafprotein between negative control group and silent group （P〉0.05）; compared with the negative con- trol group, the apoptotic rate of silent group was significantly increased, the expression of Cleaved Caspase3 protein was significantly up-regnlated, β-catenin and Cyelin D1 protein expression were down regulated （P〈0.01）. Conclusion： RNA interference of e-Maf gene could induce the apoptosis of multiple myeloma cells, which might be related to the inhibition of Wnt/β-catenin signaling pathway.