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

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.