16型人乳头状瘤病毒E6、E7诱导的人永生化口腔上皮细胞系的建立  

Establishment of human immortalized oral epithelial cell line HIO615 induced by HPV16 E6 and E7

作　　者：张志愿[1] 帕提曼.司地克 曹俊[1] 周晓健[1] 陈万涛[1] 李卿[1] 

机构地区：[1]上海第二医科大学口腔医学院口腔颌面外科,200011

出　　处：《中华口腔医学杂志》2002年第1期12-14,T003,共4页Chinese Journal of Stomatology

基　　金：国家自然科学基金资助项目 ( 39770 80 0 )

摘　　要：目的　建立 16型人乳头状瘤病毒 (humanpapiloavirus,HPV16 )E6、E7诱导的永生化口腔上皮细胞系。方法　正常口腔上皮细胞转染pLXSNHPV16E6、E7,G418筛选阳性细胞 ,连续培养建系 ;Westernblot检测细胞HPV16E6、E7蛋白表达 ;生物学特性检测 ;集落实验 ;成瘤实验。结果　细胞连续培养达 18个月余 ,命名为人永生化口腔上皮细胞 (humanimmortalizedoralepithelialcell,HIOEC) ,HIOEC表达HPV16E6、E7蛋白 ,角蛋白阳性 ,透射电镜观察细胞含有丰富的张力原纤维 ,细胞之间形成桥粒 ,克隆形成率 0 77% ,裸鼠未成瘤。结论　成功建立了HPV16E6、E7诱导的人永生化口腔上皮细胞系。Objective To establish an immortalized oral epithelial cell line Methods Normal human oral epithelial cells were transfected with HPV16E6/E7 open reading frames using recombinant retroviral system pLXSN Expression of HPV16E6 and E7 protein were tested by Western blot in three kinds of cells To define cellular biological characterization of HPV16E6/E7 transfected cells, a series analysis were performed, including protraction of growth curve, HE staining, immunocytochemical staining and scanning electron microscope observation The tumorigenicity was assessed by colony formation and transplanting the cells into nude mice Results Human oral epithelial cells transfected with HPVE6/E7 has been in culture for over 18 months The cell line was named HIO615 Western blot analysis showed HIO615 expressed HPV16 E6 and E7 protein HIOC were positive for cytokeratin, tonofibril and desmosome as observed by scanning electron microscope The number of large colonies of dense multilayer cells was low(0 77%) No tumor developed in nude mice injected subcutaneously with HIOEC Conclusion A human immortalized oral epithelial cell line induced by HPV16E6 and E7 has been successfully established

关 键 词：人乳头状瘤病毒 口腔上皮细胞 永生化 HPV16 口腔粘膜癌 

分 类 号：R739.8[医药卫生—肿瘤]