摘要
目的 观察ＥｐｓｔｅｉｎＢａｒｒ 病毒（ＥＢＶ） 对支气管上皮细胞的转化作用。方法 用ＥＢＮＡ２和ＬＭＰ１ 的真核表达质粒同时转染永生化人胚支气管上皮细胞系ＴＲ，经潮霉素Ｂ 筛选得到稳定转化细胞ＴＲ／ＬＭＰ１ＥＢＮＡ２ 。结果 原位杂交和Ｗｅｓｔｅｒｎ ｂｌｏｔ 证实ＴＲ／ＬＭＰ１ＥＢＮＡ２ 有ＥＢＮＡ２ 和ＬＭＰ１ 的ｍＲＮＡ和蛋白表达。生长曲线，ＭＴＴ显示细胞增殖能力增强，软琼脂集落形成率ＴＲ／ＬＭＰ１ＥＢＮＡ２ 为５０ ．１‰，６８ ．２‰， 而空载体转染细胞ＴＲ／Ｈｙｇ 为２．０ ‰，３．０ ‰，ＴＲ细胞为１ ．０‰，０．７‰。结论 ＬＭＰ１和ＥＢＮＡ２ 的表达使永生化支气管上皮细胞停泊依赖性降低，生长加快，细胞表型倾向更进一步转化，提示ＥＢＶ在肺癌发生中可能起一定作用。
Objective To observe Epstein Barr virus (EBV) dependent transformation of human bronchial epithelium. Methods Eukaryotic expression plasmide pBS LMP Hyg and pSG5 EBNA2 Hyg were transfected into immortalized human bronchial epithelium cell line TR.Results In situ hybridization and Western blot confirmed EBNA2 and LMP1 expression in one hygromycine B resistant cell clone TR/LMP1 EBNA2. Growth curve and MTT value showed that transformed cells grew faster. Soft agar plating rate of TR/LMP1 EBNA2 was 50.1‰, 68.2‰, empty vector PBs Hyg transfected cell strain TR/Hyg 2.0‰, 3.0‰,TR 11.0‰, 0.7‰. Conclusion Expression of both LMP1 and EBNA2 led to decreased anchorage dependence and increased proliferation rate of TR, thereby further transformed immortalized human bronchial epithelium. Our results suggest that EBV may play a role in genesis of lung cancer.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
1999年第4期277-280,共4页
Chinese Journal of Pathology
基金
中国医学科学院科研基金
关键词
肺肿瘤
上皮细胞
支气管
EB病毒
LMPI
Lung neoplasms Herpsvirus 4, human Epithelial calls Cell line, transformed
