宫颈癌HeLa细胞内HPV L1蛋白表达的观察

The expression of human papillomaviurs late capsid protein 1 in HeLa cells

目的:通过观察电镜下所见的人宫颈癌细胞株——HeLa细胞胞质内包涵体类物质的性质,了解人乳头瘤病毒(HPV)阳性宫颈癌细胞内HPV主要外壳蛋白(L1)的表达规律。方法:培养HeLa细胞,采用ELISA、光镜和电镜的免疫组织化学、Western blot方法,以HPV18 L1小鼠单克隆抗体对细胞内L1的表达情况进行检测,同时用反转录PCR方法检测细胞内L1 mRNA的存在。结果:ELISA检测显示,HeLa细胞裂解液内有HPV L1的存在。HRP标记的光镜免疫组织化学显示,培养的HeLa细胞呈现强的HPV L1阳性反应。电镜下观察,可见部分培养的HeLa细胞胞质内有团块状包涵体样物质,这些团块由大小均匀的颗粒样物质密集而成。这些颗粒性物质可为HPV L1抗体携带的胶体金颗粒标记。Western blot检测结果显示,HeLa细胞裂解液在56 kD区域,出现了一条L1特异阳性条带,说明HeLa细胞内有HPV18 L1的存在。反向PCR方法检测显示细胞内有L1 mRNA的存在。结论:电镜下所见到的培养的宫颈癌细胞株HeLa细胞胞质内的包涵体类物质,系由HPV18 L1构成,说明HeLa细胞能够表达L1。

Objective： The observations had been done to define the nature of the cytoryctes in Hela cells under electron micro- scope and to found out the regularity of HPV major capsid protein L1 express in HPV positive uterine cervix cancer cells. Methods： HPV L1 expression of cultured HeLa cells was detected by ELISA, light microscope and electron microscope immunohistochemistry as- says, Western blot assay using HPV L1 mice monoelonal antibody. L1 mRNA in HeLa cells was detected by reverse transcriptional PCR （RT-PCR）. Results：ELISA assay manifests demonstrated that HPV L1 was present in lysate of HeLa cells. HRP labeled light mi- croscope immunohistochemistry showed strong positive reaction in HeLa cells. Observed under electron microscopy, there are lumpish cytorrhyctes materials in the cytoplasm of partial HeLa cells. These cytorrhyctes materials were aggregated by very small and uniform particles and these particles can be marked by HPV L1 antibody which was labeled by colloidal gold. Western blot assay present a band at 55 kD district it was L1 specific strap which demonstrat HPV18 L1 was present in HeLa cells. Conclusion：The cytorrhyctes in HeLa cells under electron microscope are composed of HPV18 L1 and L1 can be expressed in the HeLa cells.