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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 immortalized oral epithelial cell line E6 and E7 induced by human papillomavirus type 16 (HPV16). Methods Normal oral epithelial cells were transfected into pLXSNHPV16E6, E7 and G418 to screen positive cells. The cells were cultured and cultivated continuously. Western blot was used to detect the expression of HPV16E6 and E7 proteins. The biological characteristics of the cells were detected by colony assay and tumorigenicity assay. Results The cells were cultured continuously for 18 months and named human immortalized oral epithelial cells (HIOEC). HPV16E6 and E7 proteins were expressed in HIOEC and positive for keratin. The cells were rich in fibroblasts under transmission electron microscope. Granules, colony formation rate of 0 77%, nude mice not tumor. Conclusion HPV16E6 and E7 induced human immortalized oral epithelial cell lines were established successfully.