【摘要】 目的 探讨HPV16 E7-A647G变异通过SUV39H1途径影响宫颈癌细胞的增殖和迁移。方法 宫颈癌SiHa细胞慢病毒感染HPV16 E7-647A和HPV16 E7-647G过表达载体为实验组，慢病毒空载体NC为对照组。Western blot实验检测SUV39H1、RIG-1、cGAS、STING、P-IRF3和IFN-β蛋白质表达水平；MTT实验、集落形成实验、划痕实验和Transwell实验检测HPV16 E7-A647G变异对SiHa细胞增殖及迁移能力的影响。构建HPV16 E7 siRNA干扰片段，将其转染到SiHa细胞中，检测敲低E7基因对SUV39H1通路蛋白质表达水平及细胞增殖和迁移能力的影响。结果 在宫颈癌SiHa细胞中，HPV16 E7-A647G变异通过影响SUV39H1途径蛋白质表达水平使IFN-β蛋白质表达降低，且E7-647G变异的IFN-β蛋白质表达低于E7-647A野生型，差异具有统计学意义(P<0.000 1);E7-647G变异促进宫颈癌细胞的增殖和迁移作用显著强于E7-647A野生型，差异具有统计学意义(P<0.05,P<0.01)。SiHa细胞中敲低E7基因后通过SUV39H1途径使IFN-β蛋白质表达升高，差异具有统计学意义(P<0.001),且敲低E7基因后SiHa细胞的增殖和迁移能力减弱，差异具有统计学意义(P<0.05,P<0.001,P<0.000 1)。结论 HPV16 E7-647G变异通过SUV39H1途径降低宫颈癌细胞中IFN-β的表达水平，促进宫颈癌细胞的增殖和迁移，且促进能力高于HPV16 E7-647A野生型。更多还原

【Abstract】 Objective To investigate the effect of HPV16 E7-A647G variation on the proliferation and migration of cervical cancer cells through SUV39H1 pathway. Methods Cervical cancer SiHa cell lentivirus infected with HPV16 E7-647A and HPV16 E7-647G overexpression vector as experimental group, lentivirus empty vector NC as control group. The protein expression levels of SUV39H1, RIG-1, cGAS, STING, P-IRF3 and IFN-β were detected by Western blot assay. The effects of HPV16 E7-A647G variation on SiHa cell proliferation and migration were detected by MTT assay, colony formation assay, scratch assay and transwell assay. HPV16 E7 siRNA interference fragment was constructed and transfected into SiHa cells to detect the effects of E7 knockdown on SUV39H1 pathway protein expression and cell proliferation and migration. Results In cervical cancer SiHa cells, HPV16 E7-A647G variant decreased IFN-β protein expression by affecting SUV39H1 pathway protein expression level, and the expression of IFN-β protein with E7-647G variant was lower than that of E7-647A prototype, the difference was statistically significant(P<0.000 1). The effect of E7-647G variation on the proliferation and migration of cervical cancer cells was significantly stronger than that of E7-647A prototype, and the difference was statistically significant(P<0.05, P<0.01). After E7 gene knocking down, SiHa cells increased IFN-β protein expression by affecting SUV39H1 pathway protein expression, the difference was statistically significant(P<0.001), and after E7 gene knocking down, the proliferation and migration ability of SiHa cells decreased, the difference was statistically significant(P<0.05,P<0.001, P<0.000 1). Conclusion HPV16 E7-647G variant can promote the proliferation and migration of cervical cancer cells by reducing the expression of IFN-β with SUV39H1 pathway in cervical cancer cells, and the promotion ability is higher than that of HPV16 E7-647A prototype.更多还原