重庆地区鼻咽癌患者血浆EBV-LMP1基因检测及缺失变异分析

目的:研究重庆地区鼻咽癌和非鼻咽癌患者血浆EB病毒潜伏膜蛋白l(EBV-LMP1)基因存在情况及碱基缺失变异状况,探讨其在鼻咽癌发生中的作用.方法:收集重庆籍鼻咽癌患者外周血48例,非鼻咽癌对照者外周血40例,提取DNA后,采用PCR特异性地扩增LMP1基因N端(包含Xho Ⅰ酶切位点)和C端(包含30 bp缺失的区域),N端PCR产物进行Xho Ⅰ酶切.用8%聚丙烯酰胺凝胶(PAGE)电泳分离分析酶切及PCR产物,用双脱氧终止法对部分PCR产物进行测序,用DNAssist软件对序列进行碱基缺失变异分析.结果:48例鼻咽癌外周血中,全部扩增出特异性LMP1基因条带,阳性率为100%.40例非鼻咽对照者外周血中,38例扩增出特异性条带,阳性率为95%.与B95-8原型LMP1比较,电泳分离、酶切分析、测序及软件序列分析,48例鼻咽癌患者和38例非鼻咽癌对照者外周血未发现1例存在LMP1基因N端Xho Ⅰ酶切位点和C端30 bp的缺失变异.结论:我国重庆地区鼻咽癌患者和非鼻咽癌对照者血浆携带的EBV为非缺失型(原型);LMPl基因缺失变异与鼻咽癌发病的确切关系还需进一步研究.

Analysis of EBV-LMP1 gene mutation in plasma of patients with nasopharyngeal carcinoma in chongqing

Objective:To detect and analyze the EBV-LMP1 gene N terminus XhoI-site mutation and C terminus 30bp deletion in plasma of patients with nasopharyngeal carcinoma(NPC) in Chongqing of China.Method:DNA extraction and PCR amplification was used in plasma of 48 NPC patients and 40 control non-NPC cases from Chongqing of China.All the PCR production of N terminus was digested by enzyme XhoI,then segregated in 8% PAGE.All the PCR production of C terminus was segregated in 8% PAGE too.Compared with B95-8 cell,some of the PCR production were sequenced and analyzed with software.Result:LMP1 was amplified successfully from 48 of 48 NPC cases(100%) and from 38 of 40 non-NPC cases(95%) by PCR,but none of LMP1 XhoI-site mutation and none of 30 bp deletion was found by digesting,segregating and Sequencing.Conclusion:EBV-LMP1 gene N terminus XhoI-site and C terminus 30bp deletion have no mutation in plasma of NPC or non-NPC cases in Chongqing of China.The precise relationship of EBV-LMP1 gene mutation or deletion with NPC pathogenesis needs further investigation.