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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。
Objective: To investigate the presence and deletion of Epstein-Barr virus (EBV-LMP1) gene in patients with nasopharyngeal carcinoma and non-nasopharyngeal carcinoma in Chongqing and its role in nasopharyngeal carcinogenesis. Methods: Forty-eight cases of peripheral blood from patients with nasopharyngeal carcinoma and 40 cases of peripheral blood from patients without nasopharyngeal carcinoma were collected. PCR was used to amplify the N-terminal of LMP1 gene (including Xho I restriction site) C-terminal (including 30 bp deletion region), N-terminal PCR products were digested with Xho I. The PCR products were separated by 8% polyacrylamide gel electrophoresis (PAGE), and some PCR products were sequenced by dideoxy termination method. The DNAssist software was used to analyze the base deletion mutation. Results: In 48 cases of nasopharyngeal carcinoma peripheral blood, all amplified specific LMP1 gene band, the positive rate was 100%. In 40 cases of non-nasopharyngeal controls in the peripheral blood, 38 cases amplified specific bands, the positive rate was 95%. Compared with the LMP1 B95-8 prototype, electrophoresis, enzyme digestion analysis, sequencing and software sequence analysis showed that there was no LMP1 N-terminal Xho I enzyme in 48 cases of NPC and 38 cases of non-NPC controls Cut-point and C-terminal 30 bp deletion mutation. Conclusion: EBV in plasma of patients with nasopharyngeal carcinoma and non-nasopharyngeal carcinoma in Chongqing of our country is non-deletion type (prototype). The exact relationship between LMP1 gene deletion mutation and the incidence of nasopharyngeal carcinoma needs further study.