Apak对p53R175H的调控研究

Regulation of p53R175H by Apak

目的探讨Kruppel相关盒(KRAB)型锌指蛋白Apak对p53R175H的调节功能及机制。方法报告基因检测Apak对p53R175H的活性调控,实时定量PCR检测Apak对p53R175H下游靶基因的转录水平的调控能力,Western印迹实验检测蛋白的表达。结果 Apak抑制p53175H的转录活性功能与抑制野生型p53(wtp53)的转录活性功能相比被大大减弱,Apak能够下调wtp53相关凋亡基因的水平,而在转染了p53R175H的细胞中,Apak的这种下调功能几乎丧失。结论 Apak对突变型p53R175H失去调控能力。

Objective To evaluate the regulation of p53R175H by Kruppel-associated box（KRAB）type zinc-finger protein Apak.Methods Reporter gene assay was used to detect the activity of p53 and p53R175H,and real-time fluorescent quantitation polymerase chain reaction was used to detect the level of p53 target gene.The expression level of apoptosis proteins was examined by Western blotting.Results Apak restricted the activity of wild-type p53（wtp53） but failed to regulate the activity of p53R175H,Apak negatively regulated p53-mediated target genes but had no significant effects on mutant p53-mediated target genes.Conclusion Apak cannot regulate mutant p53R175H.