人乳头瘤病毒16型E6/E7基因对人角质形成细胞增殖及其调控因子的影响

目的 研究人乳头瘤病毒（ＨＰＶ）１６型Ｅ６／Ｅ７基因对人角质形成细胞增殖及细胞周期调控因子ｐ５３,ｐＲｂ的影响。方法 Ｌｏｐｏｆｅｔａｍｉｎｅ介导法进行 染,筛选阳性克隆,免疫荧光染色检测ＨＰＶ１６Ｅ６／Ｅ７在转染细胞内的表达,流式细胞仪分析细胞周期变化,免疫印迹分析ｐ５３,ｐＲｂ表达情况。结果 免疫荧光染色检测见转染细胞内有ＨＰＶＥ６／Ｅ７表达,汉式细胞仪显示转染细胞Ｇ１期细胞为４９．２％比正常

Human papillomavirus type 16 E6/E7 affects proliferation and its regulator of keratinocytes

AIM To study the effects of human papillomavirus type 16 E6/E7 on proliferation and its regulator ofkeratinocytes.METHODS Keratinocytes were transfected with lipofectamine and the postive clone were selected by G418.The expression of HPV16E6/E7 was analysed by indirect immunofluorescent staining.Flow cytometry showed cell cycle change.The expressions of p53 and pRb were analysed by western blot. RESULTS Indirect immunofluorescent staining showed the expression of HPV16E6/E7 in transfected keratinocyte. Flow cytometry analysis revealed that normal control had a large proportion of cells in G1 phase(73 4%) and correspondingly few cells in S phase(19.4%). On the other hand, keratinocytes transfected with HPV16E6/E7 genes showed a large proportion in the S phase (39.8%)and few cells in G1 phase(49.2%). Results of Western blot showed a specific brand exhibiting a molecular weight of p53 or pRb correspondently in both normal keratinocytes and HPV16E6/E7 transfected keratinocytes. But both of p53 and pRb brand staining were less intensive in transfected keratinocytes than in normal control. CONCLUSION The HPV16E6/E7 can affect the proliferation of human keratinocytes.The p53 and pRb may play important roles in this process.