Caspase-12短发夹状RNA在内质网应激性凋亡中的作用  

Mechanism of apoptosis induced by ER stress through siRNA against caspase-12

作　　者：刘芳[1] 邹萍[1] 陈敏[2] 张敏[1] 吴耀辉[1] 肖娟[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022 [2]华中科技大学同济医学院附属协和医院泌尿外科,武汉430022

出　　处：《中华实验外科杂志》2005年第12期1568-1570,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30370595)

摘　　要：目的构建凋亡因子Caspase-12的短发夹状RNA(siRNA),观察其在体外对鼠粒单系白血病细胞WEHI-3细胞对凋亡的抵抗作用。方法设计并合成一对针对Caspase-12编码基因的siRNA,构建表达此siRNA的重组质粒psiRNA-Caspase-12,转染入WEHI-3细胞。检测胞内Cas- pase-12的mRNA和蛋白的表达。并检测转染后WEHI-3细胞的抗凋亡能力。结果酶切分析和测序证实构建成功,siRNA对Caspase-12的表达均有明显的抑制作用。染后WEHI-3细胞的抗凋亡能力增加。结论构建的重组质粒能有效抑制Caspase-12基因在白血病细胞株WEHI-3中的表达。并对内质网应激性凋亡具有抵抗能力。Objective To silence caspase-12 expression in WEHI-3 cells using vector-based RNA interference （RNAi） technique and investigate the anti-apoptotic effect of siRNA against caspase-12 in mouse leukemia cell WEHI-3, Methods The small inhibitory RNA （siRNA） used to silence caspase-12 was designed and synthesized. The recombinant plasmid （psiRNA-caspase-12） expressing slRNA was constructed and transfected into the WEHI-3 cells, The expression level of caspase12 was examined by RT-PCR and Western blot, and the resistive effect of siRNA on apoptosis was examined. Results The recombinant plasmid psiRNA-caspase-12 was successfully construc＇ced. The recombinant plasmid suppressed the caspase-12 expression significantly. Conclusion Caspase-12 expression in WEHI-3 cells can be inhibited significantly by plasmid based RNAi. The further study demonstrated plasmld based RNAi plasmid based RNAi possessed resistive ability in apoptosis initiated by endoplasmic reticulum stress.

关 键 词：RNA干扰 CASPASE-12 益康唑 内质网 应激性凋亡 白血病 

分 类 号：R363[医药卫生—病理学]