EB病毒LMP1促鼻咽上皮细胞增殖的蛋白分子鉴定  

Identification of proteins related to proliferation and regulated by Epstein Barr virus encoded latent membrane protein 1 in nasopharyngeal epithelail cells

作　　者：张志伟[1,2,3] 张琼[1] 刘洁琼[1] 余艳辉[1] 贺智敏[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所,湖南长沙410078 [2]南华大学医学院肿瘤研究所 [3]肿瘤细胞与分子病理学湖南省高校重点实验室,湖南衡阳421001

出　　处：《中国病理生理杂志》2010年第2期287-292,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30470668);湖南省自然科学基金资助项目(No.07JJ6164)

摘　　要：目的:探讨EB病毒潜伏性膜蛋白1(LMP1)促鼻咽上皮细胞增殖的分子机制。方法:采用逆病毒感染的方法,将浓缩的逆病毒RV-pLNSX(空载体)和RV-LMP1分别感染鼻咽上皮细胞NP69,建立NP69-pLN-SX与NP69-LMP1稳定表达细胞系,通过绘制细胞生长曲线、平皿克隆形成实验和软琼脂集落形成实验检测LMP1对NP69细胞增殖的影响;运用比较蛋白质组学方法鉴定LMP1在NP69细胞中参与调节的蛋白,并对部分蛋白表达进行验证。结果:(1)LMP1具有促鼻咽上皮细胞NP69增殖的作用(n=3,P<0.05)。(2)鉴定了22个LMP1参与调节NP69细胞的蛋白(表达上调的蛋白9个,下调的13个),实时荧光定量RT-PCR和Western blotting证实了部分上述蛋白的差异表达。结论:LMP1可能通过参与调节keratin19和vimentin等蛋白的表达促鼻咽上皮细胞NP69增殖。AIM ： To investigate the molecular mechanism of Epstein - Barr virus encoded latent membrane protein 1 regulated cellular proliferation in nasopharyngeal epithelial cells. METHODS： The nasopharyngeal epithelial cells NP69 were infected with RV -pLNSX （the empty vector） and RV- LMP1 retroviruses, respectively. Therefore, the NP69 - pLNSX and NP69 - LMP1 cell lines were established. Sequentially, cellular proliferation of NP69 - pLNSX and NP69 - LMP1 cells was compared to draw the cellular growth curve. The experiments of plate clone formation and forming of soft agar colony were conducted. Meanwhile, the differential expression of proteins were identified between NP69 - pLN- SX and NP69 - LMP1 cell lines by proteomie methods, and the expression levels of partial identified proteins were verified. RESULTS ： （ 1 ） LMP1 was able to accelerate cellular proliferation of nasopharyngeal epithelial cell NP69 （ n = 3, P 〈 0. 05）. （2） Twenty two proteins （9 up -and 13 down -regulated） of LMP1 mediated regulation were identified from in- fected NP69 cell lines, and the differential expression of partial identified proteins was confirmed by Western blotting and fluorescent real - time quantitative RT - PCR. CONCLUSION： LMP1 probably mediates the regulation of vimentin protein and keratin 19 protein expression to promote cellular proliferation in NP69 cells.

关 键 词：EB病毒 潜伏性膜蛋白1 细胞增殖 差异蛋白表达 

分 类 号：R318.0[医药卫生—生物医学工程]