【作者】 张志愿； 帕提曼； 曹俊； 周晓健； 李卿； 陈万涛；

【Author】 Zhang Zhiyuan Sdek Patima Cao Jun Zhou Xiaojian Li Qing Chen Wantao Dept of Oral & Maxillofacial Surgery,Shanghai Ninth People’s Hospital, Shanghai Second Medical University

【机构】 上海第二医科大学附属第九人民医院；

【摘要】 <正>目的:建立16型人乳头状瘤病毒(HPV16)E6、E7诱导的永生化口腔上皮细胞系。材料与方法: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:Westrn blot检测细胞HPV16E6、E7蛋白表达;细胞生物学特性检测;集落试验;成瘤试验。结果:细胞连续培养18更多还原

【Abstract】 Objective: To establish an immortalized oral epithelial cell line. Materials and Methods:Normal human oral epithelial cells were transfected with HPV16E/E7 open reading frames using redcombinant 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 HPV16E/E7 transfected cells, a series analysis were performed, include protraction of growth curve, HE staining,immunocytochemical staining and scanning electron microscope observation. Soft agar culture and subcutaneous injection of HIOEC from culture into athymic nude mice. Results: Human oral epithelial cells transfected with HPVE6/E7 have been in culture for veor 18 months. The cell line was named human immortalized oral epithelial cell (HIOEC). Western blot analysis showed HIOEC expressed HPV 16 E6 and E7 protein. HIOEC lost contact inhibition. The doublet proliferation time is 54.37 hours. were positive for cytokeratin, tonofibril and demosome can be observed by scanning electron microscope. Cloning efficiency is about 7.7%. No neoplasm was detected in athymic nude mice. Conclusion: A human immortalized oral epithelial cell line indued by HPV16E6 and E7 has been successfully established.更多还原