基础医学与临床
基礎醫學與臨床
기출의학여림상
Basic & Clinical Medicine
2015年
1期
60-64
,共5页
杜义江%肖长义%郑军%胡敏
杜義江%肖長義%鄭軍%鬍敏
두의강%초장의%정군%호민
HepG2细胞%人乳头瘤病毒%病毒基因组%物理状态%L1蛋白
HepG2細胞%人乳頭瘤病毒%病毒基因組%物理狀態%L1蛋白
HepG2세포%인유두류병독%병독기인조%물리상태%L1단백
HepG2 cells%human papillomavirus%virus genome%physical state%late capsid protein 1
目的：了解人肝癌细胞系HepG2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HepG2细胞内L1蛋白表达；用反转录PCR检测细胞内L1 mRNA表达。结果 HepG2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。 HepG2细胞裂解液中有HPV L1蛋白，在56 ku区出现与HeLa细胞一样的L1特异阳性条带。反转录PCR检测显示HepG2细胞内有L1 mRNA存在。结论 HepG2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，HepG2细胞可以表达L1。
目的：瞭解人肝癌細胞繫HepG2細胞內人乳頭瘤病毒（ HPV）基因組的物理狀態，胞質內包涵體物質的性質以及晚期衣殼蛋白1（L1）錶達。方法用PCR對細胞內HPV18型E2和E6基因進行擴增，判斷HPV18基因組的物理狀態；用ELISA、光鏡和電鏡的免疫組化、Western blot 等方法，以多價HPV L1小鼠單剋隆抗體做探針，檢測HepG2細胞內L1蛋白錶達；用反轉錄PCR檢測細胞內L1 mRNA錶達。結果 HepG2細胞內HPV DNA基因組呈整閤狀態；細胞裂解液中有HPV L1蛋白存在；細胞呈HPV L1暘性反應；胞質內包涵體樣物質，由均勻的顆粒樣物質組成，可以為膠體金標記的HPV L1抗體所標識。 HepG2細胞裂解液中有HPV L1蛋白，在56 ku區齣現與HeLa細胞一樣的L1特異暘性條帶。反轉錄PCR檢測顯示HepG2細胞內有L1 mRNA存在。結論 HepG2是HPV18暘性細胞，細胞內HPV DNA基因組呈整閤狀態。細胞內包涵體樣物質為HPV18 L1蛋白，HepG2細胞可以錶達L1。
목적：료해인간암세포계HepG2세포내인유두류병독（ HPV）기인조적물리상태，포질내포함체물질적성질이급만기의각단백1（L1）표체。방법용PCR대세포내HPV18형E2화E6기인진행확증，판단HPV18기인조적물리상태；용ELISA、광경화전경적면역조화、Western blot 등방법，이다개HPV L1소서단극륭항체주탐침，검측HepG2세포내L1단백표체；용반전록PCR검측세포내L1 mRNA표체。결과 HepG2세포내HPV DNA기인조정정합상태；세포렬해액중유HPV L1단백존재；세포정HPV L1양성반응；포질내포함체양물질，유균균적과립양물질조성，가이위효체금표기적HPV L1항체소표식。 HepG2세포렬해액중유HPV L1단백，재56 ku구출현여HeLa세포일양적L1특이양성조대。반전록PCR검측현시HepG2세포내유L1 mRNA존재。결론 HepG2시HPV18양성세포，세포내HPV DNA기인조정정합상태。세포내포함체양물질위HPV18 L1단백，HepG2세포가이표체L1。
Objective To find out the physical state of the human papillomavirus ( HPV) genome in hepatoma cell line HepG2 cells and the regulation of HPV late capsid protein 1 ( L1) expression and to explore the nature of the cytoryctes in HepG2 cells.Methods E2 and E6 in HPV18 were detected by PCR to evaluate the physical state of HPV18 genome .HepG2 L1 expression was detected by ELISA , light microscropy and electron microscrope immu-nohistochemistry assays , Western blot assay using HPV L 1 mice monoclonal antibody .L1 mRNA in HepG2 cells was detected by reverse transcriptional PCR ( RT-PCR) .Results PCR assay displayed that HPV DNA was inte-grated with HepG2 genome.ELISA assay showed that HPV L1 was present in lysate of HepG2 cells.Light micros-cropy demonstrated strong positive reaction in HepG2 cells.In microscopy, in the cytoplasm of partial HepG2 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 dis-trict and it was L1 specific strap which demonstrated HPV 18 L1 was present in HepG2 cells.RT-PCR assay demon-strated the presence of L1 mRNA in HepG2 cells.Conclusions HepG2 cells are HPV18-positive HPV DNA ge-nome is integrated with HepG2 cells.HepG2 cells can express L1.The cytorrhyctes in HepG2 cells are composed of HPV18 L1 indicating that L1 can be expressed in HepG2.
