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目的 研究 16型人乳头瘤病毒 (HPV ) E6 / E7对人角质形成细胞细胞周期素 (Cyclin A,Cyclin D1 )及细胞周期素依赖性激素 (cdk4)的影响 .方法 用 L ipofectamine介导法将p L XSN16 E6 / E7质粒转染逆转录病毒包装细胞 PA317,挑选G418抗性克隆 ,检测病毒滴度 ,将表达 HPV16 E6 / E7基因的高滴度逆转录病毒感染人角质形成细胞 ;免疫印迹分析感染细胞 Cyclin A,Cyclin D1 ,cdk4表达情况 .结果 感染表达HPV16 E6 / E7基因的逆转录病毒的角质形成细胞生长时间明显延长 ,免疫印迹分析发现 ,感染细胞 Cyclin A和 cdk4表达增多 ,而 Cyclin D1 表达明显减少 .结论 HPV16 E6 / E7能影响细胞的生长和增殖 ,同时伴有细胞周期调节蛋白 Cyclin A,Cyclin D1 及 cdk4的表达异常 .这些细胞周期蛋白表达异常发生于转染早期 ,也许是细胞逃脱衰老 ,开始永生化所必需的
Objective To investigate the effects of HPV E6 / E7 on the expression of Cyclin A (Cyclin D1) and cyclin-dependent hormone (cdk4) in human keratinocytes.Methods The p16 L XSN16 E6 / E7 plasmid was transfected into retroviral packaging cell line PA317, G418-resistant clones were selected to detect the virus titer, high-titre retrovirus infected with HPV16 E6 / E7 gene was used to infect human keratinocytes, and immunoblotting was used to analyze the infection The expression of Cyclin A, Cyclin D1 and cdk4 were detected by Western blotting.Results The growth of keratinocytes infected with retrovirus expressing HPV16 E6 / E7 gene was significantly prolonged, and the expression of Cyclin A and cdk4 in infected cells was increased by Western blot analysis, while the expression of Cyclin D1 Significantly decreased.Conclusion HPV16 E6 / E7 can affect the cell growth and proliferation, accompanied by the abnormal expression of cell cycle regulatory proteins Cyclin A, Cyclin D1 and cdk4.The abnormal expression of these cell cycle proteins occurred in the early stage of transfection, which may be cell escape Aging, necessary to begin immortality