摘要
目的了解人肝癌细胞系Hep G2细胞内人乳头瘤病毒(HPV)基因组的物理状态,胞质内包涵体物质的性质以及晚期衣壳蛋白1(L1)表达。方法用PCR对细胞内HPV18型E2和E6基因进行扩增,判断HPV18基因组的物理状态;用ELISA、光镜和电镜的免疫组化、Western blot等方法,以多价HPV L1小鼠单克隆抗体做探针,检测Hep G2细胞内L1蛋白表达;用反转录PCR检测细胞内L1 mRNA表达。结果 Hep G2细胞内HPV DNA基因组呈整合状态;细胞裂解液中有HPV L1蛋白存在;细胞呈HPV L1阳性反应;胞质内包涵体样物质,由均匀的颗粒样物质组成,可以为胶体金标记的HPV L1抗体所标识。Hep G2细胞裂解液中有HPV L1蛋白,在56 ku区出现与He La细胞一样的L1特异阳性条带。反转录PCR检测显示Hep G2细胞内有L1 mRNA存在。结论 Hep G2是HPV18阳性细胞,细胞内HPV DNA基因组呈整合状态。细胞内包涵体样物质为HPV18 L1蛋白,Hep G2细胞可以表达L1。
Objective To find out the physical state of the human papillomavirus( HPV) genome in hepatoma cell line Hep G2 cells and the regulation of HPV late capsid protein 1( L1) expression and to explore the nature of the cytoryctes in Hep G2 cells. Methods E2 and E6 in HPV18 were detected by PCR to evaluate the physical state of HPV18 genome. Hep G2 L1 expression was detected by ELISA,light microscropy and electron microscrope immunohistochemistry assays,Western blot assay using HPV L1 mice monoclonal antibody. L1 mRNA in Hep G2 cells was detected by reverse transcriptional PCR( RT-PCR). Results PCR assay displayed that HPV DNA was integrated with Hep G2 genome. ELISA assay showed that HPV L1 was present in lysate of Hep G2 cells. Light microscropy demonstrated strong positive reaction in Hep G2 cells. In microscopy,in the cytoplasm of partial Hep G2 cells,there were lumpish cytorrhyctes materials which consists of very small and uniform particles and these parti-cles were marked by HPV L1 antibody labeled by colloidal gold. Western blot analysis showed a band at 56 ku district and it was L1 specific strap which demonstrated HPV18 L1 was present in Hep G2 cells. RT-PCR assay demonstrated the presence of L1 mRNA in Hep G2 cells. Conclusions Hep G2 cells are HPV18-positive HPV DNA genome is integrated with Hep G2 cells. Hep G2 cells can express L1. The cytorrhyctes in Hep G2 cells are composed of HPV18 L1 indicating that L1 can be expressed in Hep G2.
出处
《基础医学与临床》
CSCD
2015年第1期60-64,共5页
Basic and Clinical Medicine
基金
湖北省自然科学基金(2011CDC002)
