HPV16E6对Hippo途径的效应分子YAP的影响

The Effect of HPV16E6 Protein on Effector Molecule YAP of the Hippo Pathway

目的研究HPV16E6和Hippo途径的效应分子YAP的相关性。方法用Western Blot、Real-Time qRTPCR和激光扫描共聚焦显微镜观察Caski细胞HPV16E6过表达或被敲减后YAP的表达和定位及其直接作用的下游基因结缔组织生长因子(CTGF)的表达。结果 Caski细胞HPV16E6被敲减或过表达时,YAP相应地表达减少或增加,磷酸化YAP增加或减少,YAP下游分子CTGF的mRNA表达相对应减少或升高。YAP主要定位于胞浆或胞核胞浆中,磷酸化YAP主要定位于胞浆,非磷酸化YAP主要定位于胞核。结论 HPV16E6诱导Hippo途径失常,胞核YAP表达升高,CTGF表达相应增多,提升HPV16E6的致癌性。Hippo途径可能是治疗高危性HPV16感染相关疾病的候选药物作用途径之一。

Objective To explore the correlation between the effect of HPV16E6 and the molecular YAP of Hippo pathway. Methods The expression and localization of YAP and its downstream connective tissue growth factor（ CTGF） were analyzed by using Western blot,Real-Time qRT-PCR or laser scanning confocal microscope after HPV16E6 overexpression or knockdown in the Caski cell. Results When HPV16E6 was knocked down or over expressed,the corresponding expression of YAP or phosphorylation of YAP decreased or increased. The mRNA expression of CTGF,a YAP downstream molecule,aslo decreased or increased correspondly. The phosphorylation of YAP was mainly localized in the cytoplasm,and the non phosphorylated YAP was mainly localized in the nucleus. Conclusion HPV16E6 induces Hippo pathway deregulation,YAP overexpression and CTGF mRNA expression. HPV16E6 induced disorders of Hippo pathway and increased nuclear YAP expression,which enhanced CTGF expression and promoted the carcinogenicity of HPV16E6. They might promote HPV16E6 pathogenesis,implying that Hippo pathway might be candidate drug target of hr-HPV infection-induced diseases. Hippo pathway may be one of the candidate pathway for the drug treatment of high risk HPV16 infection related diseases