Bcr/abl融合基因的小干扰RNA对K562细胞增殖和凋亡的影响  

Effect of bcr/abl fusion gene siRNA on proliferation and apoptosis of K562 cells

作　　者：张小鹰[1] 曾慧兰[2] 蒋建伟[1] 陈涛[1] 吴风云[2] 何金花[1] 韩新爱[2] 廖晓莉[1] 

机构地区：[1]暨南大学医学院生物化学教研室,广州510630 [2]暨南大学附属第一医院,广州510632

出　　处：《广东医学》2008年第11期1772-1775,共4页Guangdong Medical Journal

基　　金：广东省自然科学基金博士启动基金项目(编号:06300580);广东省医学科学技术研究基金项目(编号:A2006352);广东省中医药管理局基金项目(编号:1040068)

摘　　要：目的观察特异性bcr/abl融合基因的siRNA对慢性粒细胞白血病K562细胞增殖和凋亡的影响。方法设计并化学合成bcr/abl融合基因融合位点b3:a221个核苷酸siRNA作用于K562细胞,用WST-8法检测细胞增殖抑制率;RT-PCR检测bcr/abl mRNA表达水平;PI单染流式细胞仪检测细胞周期;AnnexinV-PI双染测定细胞凋亡比例;Hochest33258染色荧光显微镜观察细胞凋亡的形态学变化。结果①Bcr/ablsiRNA作用K562细胞24,48,72h后,明显抑制K562细胞增殖,各浓度组间的增殖抑制率差异无显著性(P>0.05);②Bcr/ablsiRNA能显著下调bcr/abl mRNA水平,各浓度组间差异无显著性(P>0.05);③Bcr/ablsiRNA作用组细胞周期阻滞于G1期;④Bcr/ablsiRNA作用后细胞出现明显的早期凋亡群,各浓度组间的早期凋亡率差异无统计学意义(P>0.05),细胞出现核固缩、核边集、凋亡小体等改变。结论特异性bcr/ablsiRNA可显着抑制K562细胞bcr/abl融合基因的表达,抑制细胞的增殖,诱导凋亡,但其作用未显示明显的剂量依赖性。Objective To investigate the effect of bcr/abl fusion gene small interfering RNA （siRNA）on the proliferation and apoptosis of chronic myelogenous leukemia （CML） K562 cells. Methods A 21nt siRNA targeting b3 ： a2 mRNA of bcr/abl fusion gene was designed, synthesized, and transfected into K562 cells. The proliferation inhibition rate was measured by WST- 8 method; The expression of bcr/abl mRNA of K562 cells was detected by RT- PCR; The cell cycle was determined by Phosphatidylinositol （PI） staining; The apoptosis rate was detected by Annexin V - fluorescencein isothiocyanate/phosphatidylinositol （PI） double staining; The morphology of apoptosis was examined by Hochest 33258 staining. Results ①Bcr/abl siRNA significantly inhibited K562 cells proliferation 24 h, 48 h and 72 h after transfection. ②Bcr/abl siRNA significantly down - regulated mRNA expression level. There was no significant difference between among different concentration bcr/abl siRNA groups （P 〉 0.05）. ③Cell cycle of K562 cells was blocked in G1 phase after bcr/abl siRNA transfection. ④Apoptotic cells appeared early after bcr/abl siRNA transfection. The early apoptotic rates were not significantly different among different concentration bcr/abl siRNA groups （ P 〉 0.05 ）. Cells presented karyopyknoses, nuclear - set and apoptotic bodies. Conclusion The specific bcr/abl fusion gene siRNA significantly inhibits the expression of bcr/abl fusion gene, inhibits K562 cell proliferation and induces its apoptosis, but the efficacy was not dose - dependent.

关 键 词：BCR/ABL SIRNA K562细胞 增殖 凋亡 

分 类 号：R733.72[医药卫生—肿瘤] R971.1[医药卫生—临床医学]