HIV感染者体内Epstein-Barr病毒致癌性潜伏膜蛋白-1功能区C末端DNA序列分析  

DNA sequence analysis of functional C-terminus of carcinogeneous latent membrane protein-1 of Epstein-Barr virus in HIV-infected patients

作　　者：周素娟[1] 丁盈盈[1] 高眉扬[1] 陈莉[1] 张铁军[1] 何纳[1] 

机构地区：[1]复旦大学公共卫生学院流行病学教研室-公共卫生安全教育部重点实验室,上海200032

出　　处：《复旦学报（医学版）》2011年第6期518-521,526,共5页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金资助项目(30671880);上海市教委科研创新项目(08ZZ02)

摘　　要：目的初步了解在HIV感染者体内,Epstein-Barr病毒(Epstein-Barr virus,EBV)致癌性潜伏膜蛋白-1(latent membrane protain-1,LMP-1)功能区C末端DNA特异性30bp缺失的发生率和33bp重复序列拷贝数的情况。方法采用巢式PCR扩增175例HIV感染者外周血淋巴细胞(peripheral blood lymphocyte,PBL)中EBV的LMP-1功能区C末端的基因片段,并进行核酸序列分析。为了避免核酸质量对实验成功率的影响,同时对提取的所有标本DNA的MDR1 C3435 T基因进行PCR扩增。结果所有HIV感染者的PBL标本中,MDR1 C3435 T基因均成功扩增(扩增率为100%),证明标本的DNA质量合格。175例标本中的33例可以成功扩增EBV-DNA(阳性率为18.9%)。在33例EBV-DNA阳性的标本中,27例(81.8%)的LMP-1基因核酸序列发生特异性30bp缺失(the 30bp-del),其33bp重复序列的拷贝数分别为4(4例,14.8%)、5(12例,44.4%)、6(8例,29.6%)、7(3例,11.1%);其他6例LMP-1核酸序列没有发生30bp核甘酸缺失,其33bp重复序列单位的拷贝数为4(5例,83.3%)、5(1例,16.7%)。结论 HIV感染者体内EBV致癌性LMP-1功能区C末端基因发生特异性30bp缺失的频率为81.8%,而其33bp重复序列的拷贝数为4、5、6、7,共4种。Objective To explore the specific 30 bp deletion（the 30 bp-del） rate and copies of 33 bp repeats of functional C-terminus of carcinogeneous latent membrane protein-1（LMP-1） of Epstein-Barr virus（EBV） in HIV-infected patients.Methods The functional C-terminal region of LMP-1 gene was amplified from peripheral blood lymphocyte（PBL） of 175 HIV infected patients by nest-PCR,and was further subject to DNA sequence analysis.To ensure the quality of extracted DNA,MDR1C3435T gene was amplified for each sample at the same time.Results All the 175 samples could successfully amplify MDR1C3435T gene by PCR.Among the 175 subjects,33（18.9%） were successfully amplified with EBV-DNA.A mong them,27 cases were were found to have the 30 bp-del in their LMP-1 gene,and had 4（4 cases,14.8%）,5（12 cases,44.4%）,6（8 cases,29.6%）,or 7（3 cases,11.1%）copies of the 33 bp DNA repeats,respectively,in their LMP-1 gene.The 30 bp-del in LMP-1 gene was absent for the other 6 subjects,of whom the 33 bp DNA repeats occurred for 4（5 cases,83.3%） or 5（1 case,16.7%）copies,respectively.Conclusions In the gene of the functional C-ter minus of carcinogeneous LMP-1 of EBV in HIV-infected patients,the frequency of the 30 bp-del was 81.8%,and had 4 kinds of copy numbers of the 33 bp repeats.

关 键 词：HIV感染者 EPSTEIN-BARR病毒 潜伏膜蛋白-1 巢式PCR 

分 类 号：R512.91[医药卫生—内科学]