RNAi沉默XIAP基因抑制MG-63细胞增殖及其分子机制  

Inhibitory effect of XIAP gene silencing induced by RNA interference on MG-63 cells and its mechanism

作　　者：何畔[1] 梁珊[2] 王愉思[1] 魏磊平[1] 阎戈[1] 康亦峰[1] 王靖[1] 

机构地区：[1]湖南省人民医院骨科,长沙市410005 [2]湖南省人民医院临床医学研究所,长沙市410005

出　　处：《江苏医药》2011年第2期148-151,共4页Jiangsu Medical Journal

摘　　要：目的探讨RNA干扰(RNAi)抑制MG-63骨肉瘤细胞X-连锁凋亡抑制蛋白(XIAP)基因的表达及其抗肿瘤的机制。方法构建XIAP基因的短发夹RNA(shRNA)表达载体。将MG-63细胞分为空白组(A组),用空白质粒psiRNA-Con转染(B组),用psiRNA-XIAP转染(C组)。Western blot检测XIAP蛋白的表达;MTT检测细胞的增殖;流式细胞术检测细胞周期、凋亡及半胱天冬氨酸蛋白酶9(Caspase-9)的表达;ELISA法检测细胞内血管内皮生长因子(VEGF)含量。结果与A、B组比较,C组细胞XIAP蛋白水平、MTT值及VEGF含量均显著降低,细胞凋亡率、Caspase-9蛋白表达及G2/M期细胞所占比例增高(P<0.05)。结论 RNAi能有效抑制XIAP基因表达,抑制骨肉瘤细胞增殖活性,诱导其凋亡。其机制可能涉及Caspase家族凋亡信号转导通路的活化、DNA合成及肿瘤血管生成的抑制。Objective To investigate the inhibitory effect of RNA interferece （RNAi） on the expression of X-linked inhibitor of apoptosis protein（XIAP） gene in MG-63 osteosarcoma cells and its anti-tumor mechanism.Methods The plasmid for expressing short hairpin RNA（shRNA） targeting XIAP gene was constructed.MG-63 cells were conventionally cultured and divided into groups of A（non-transfenction）,B（blank plasmid psiRNA-Con transfencted） and C（psiRNA-XIAP transfected）.XIAP expression in MG-63 cells was detected by Western blot.The proliferation activity in vitro was examined by MTT assay.Flow cytometry was employed to detect the percentage of cells in each phase of the cell cycle,cell apoptosis and Caspase-9 expression.The level of vascular endothelial growth factor（VEGF） was examined by ELISA.Results Compared to groups of A and B,the levels of XIAP,VEGF and MTT value were lower,apoptosis,Caspase-9 expression and G2/M cell cycle arrest were higher in group C（P0.05）.Conclusion RNAi can specifically suppress XIAP gene expression,effectively inhibit the proliferation and accelerate the apoptosis of MG-63 cell line,which may be related to activation of Caspase-9 signaling pathway,inhibition of DNA synthesis and tumor angiogenesis.

关 键 词：RNA干扰 X-连锁凋亡抑制蛋白 骨肉瘤 

分 类 号：R738[医药卫生—肿瘤]