人乳头瘤病毒16型E6/E7基因对人角质形成细胞生长及CyclinA,CyclinD1和cdk4的影响

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（ＣｙｃｌｉｎＡ，ＣｙｃｌｉｎＤ１）及细胞周期素依赖（ｃｄｋ４）的影响。方法 用Ｌｉｐｏｆｅｃｔａｍｉｎｅ介导法将ＰＬＸＳＮ１６Ｅ６／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原高滴度逆转 录病毒感染人角质形成细胞；免疫印迹分析感染细胞ＣｙｃｌｉｎＡ，ＣｙｃｌｉｎＤ１，ｃｄ

Influence of human papillomavirus type16 E6 and E7 gene on keratinocyte growth and the expression of CyclinA,CyclinD1 and cdk4

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.