人乳头瘤病毒16型E6/E7基因对人角质形成细胞生长及CyclinA,CyclinD_1和cdk4的影响  

Influence of human papillomavirus type16 E6 and E7 gene on keratinocyte growth and the expression of CyclinA, CyclinD 1 and cdk4

作　　者：马翠玲[1] 刘玉峰[1] 李志强[2] 廖文俊[1] 

机构地区：[1]第四军医大学西京医院皮肤科,陕西西安710033 [2]第三军医大学西南医院皮肤科

出　　处：《第四军医大学学报》2000年第9期1156-1158,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金!(395 70 6 5 6 )

摘　　要：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 。AIM To study the influence of human papillomavirus (HPV) type16 E6 and E7 gene on keratinocyte growth and the expression of CyclinA, CyclinD 1 and (Cyclin dependent kinases, cdk)4. METHODS The pLXSN16E6/E7 were first transfected into the retrovirus packaging cell line PA317 using the lipofectamine transduction. The G418 resistant colonies were picked and assayed for virus titration. The human keratinocytes were then infected with these retrovirus expressing the HPV16 E6/E7 genes. The expression of CyclinA, CyclinD 1 and cdk4 was analysed by western blot. RESULTS The human keratinocytes infected with the retrovirus expressing the HPV16 E6/E7 genes showed prolonged growth potential. Results of western blot suggested the HPVE6/E7 transducted cells expressed high levels of CyclinA and cdk4 but lower levels of Cyclin D 1. CONCLUSION The HPVE6/E7 can affect the growth and proloferation of human keratinocytes, and may be accompanied with the discordant exprssion of cell regulatory proteins Cycline A, Cycline D 1 and cdk4. These discordant expressions of cell regulatory proteins are early events which may be necessary for cells to escape senescence and initiate immortalization.

关 键 词：人乳头瘤病毒 角蛋白细胞 细胞周期蛋白 

分 类 号：R329-28[医药卫生—人体解剖和组织胚胎学] R373[医药卫生—基础医学]