HPV16癌基因调节STAT3磷酸化和miR-21的表达

HPV16 oncogenes regulate the phosphorylation of STAT3 and expression of miR-21.

目的:研究人乳头瘤病毒16型(HPV16)癌基因(E6和E7)与信号传导与转录激活因子3(STAT3)和微RNA-21(miR-21)表达的关系。方法:应用RNA干扰(RNAi)沉默HPV16阳性的宫颈癌细胞株SiHa细胞中HPV16的E6和E7,应用慢病毒表达载体在HPV阴性的角质形成细胞HaCaT中表达HPV16 E6和E7蛋白。Western blot法检测STAT3总蛋白的表达和磷酸化(Y705和S727位点)水平,实时定量逆转录聚合酶链反应(QRT-PCR)检测miR-21的表达。结果:沉默SiHa细胞中HPV16 E6和E7基因后,STAT3的磷酸化水平和miR-21的表达降低,STAT3的总蛋白水平无明显变化;在HaCaT细胞中表达HPV16 E6和E7蛋白后,STAT3的磷酸化水平和miR-21表达升高,STAT3的总蛋白水平无明显变化。结论:本研究首次表明HPV16的癌基因能够调节STAT3的磷酸化和miR-21的表达,提示STAT3或miR-21可能成为治疗宫颈癌及其癌前病变的分子靶点。

Objective： To investigate the relationship among human papillomavirus type 16（HPV 16） oncogenes,signal transducer and activator of transcription 3（STAT3） and miR21.Methods： HPV16 oncogenes E6 and E7 were either silenced by RNA interference（RNAi） in HPV16 positive cervical cancer cell line SiHa cells,or expressed in HPV 16 negative keratinocytes HaCaT cells.STAT3 expression and its phosphorylation at Y705 and S727 residues were analyzed by Western blot and expression of miR-21 was analyzed by quantitative reverse transcription polymerase chain reaction（QRT-PCR）.Result： Phosphorylation of STAT3 and expression of miR-21 decreased after silence of HPV16 E6 and E7 genes in SiHa cells and increased after expression of E6 and E7 proteins in HaCaT cells,but the expression of STAT3 protein was not changed significantly.Conclusion： This study demonstrates for the first time that both the phosphorylation of STAT3 and expression of miR-21 are regulated by HPV16 oncogenes.STAT3 or miR-21 could be a molecular target for treatment of cervical cancer and precancerous lesions.