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背景与目的;众所周知,EB病毒LMP1基因在鼻咽癌变过程起着一定的作用。本研究通过检测广东地区鼻咽癌组织EB病毒LMP1基因N-末端区Xho Ⅰ酶切位点的丢失,探讨LMP1基因变异在鼻咽癌发生发展中的作用。方法:收集中山大学肿瘤防治中心鼻咽癌患者鼻咽新鲜活检标本63例。收集EB病毒健康携带者外周血单个核细胞(PBMCs)10例作为对照。采用QIAamp DNA Mini Kit和QIAampDNA Blood Mini Kit分别抽取组织和外周血单个核细胞的DNA,应用巢式PCR扩增EB病毒LMP1基因的N-末端区,并用Xho Ⅰ对扩增产物进行酶切。采用四色荧光 末端终止法对扩增产物进行序列分析。结果:10例健康携带者外周血单个核细胞的EB病毒LMP1基因N-末端区均未见Xho Ⅰ酶切位点的丢失。63例鼻咽癌组织中有50例(79.37％)出现Xho Ⅰ酶切位点的丢失(Xho Ⅰ-loss),还有4例(6.34％)为Xho Ⅰ酶切位点部分丢失,只有9例(14.29％)未见Xho Ⅰ酶切位点的丢失(wt-Xho Ⅰ)。除了Xho Ⅰ酶切位点的丢失(nt:169423～169428;GAGCTC→GA T CTC)外,还发现四个错义点突变。结论:本研究所检测的广东地区EB病毒健康携带者外周血单个核细胞所携带的EB病毒LMP1基因为wt-Xho Ⅰ,而在鼻咽癌组织中主要为Xho-Ⅰ-loss。因此,我们认为EB病毒LMP1基因N-末端区Xho Ⅰ酶切位点的丢失和其?
Background and purpose; As we all know, Epstein-Barr virus LMP1 gene plays a role in the process of nasopharyngeal carcinogenesis. In this study, the detection of LMP1 gene mutation in the occurrence and development of nasopharyngeal carcinoma by detecting the loss of Xho Ⅰ restriction site in the N-terminal region of LMP1 gene of nasopharyngeal carcinoma in Guangdong Province. Methods: 63 cases of nasopharyngeal fresh biopsy specimens from patients with nasopharyngeal carcinoma in Sun Yat-sen University Cancer Center were collected. Peripheral blood mononuclear cells (PBMCs) from 10 EB virus healthy carriers were collected as control. The QIAamp DNA Mini Kit and QIAamp DNA Blood Mini Kit were used to extract DNA from tissue and peripheral blood mononuclear cells. The N-terminal region of EBV LMP1 gene was amplified by nested PCR. The amplified product was digested with Xho Ⅰ. The amplification products were sequenced by four-color fluorescence termination. Results: No deletion of Xho Ⅰ restriction site was found in the N-terminal region of EBV LMP1 gene in peripheral blood mononuclear cells from 10 healthy donors. Xho Ⅰ-loss was found in 50 cases (79.37%) of 63 cases of nasopharyngeal carcinoma, and 4 cases (6.34%) were partly lost in Xho Ⅰ restriction site, only 9 Cases (14.29%) did not see Xho Ⅰ enzyme site loss (wt-Xho Ⅰ). In addition to the loss of Xho I restriction sites (nt: 169423-169428; GAGCTC → GA T CTC), four missense point mutations were also found. Conclusion: The Epstein-Barr virus LMP1 gene carried by peripheral blood mononuclear cells of healthy carriers of Epstein-Barr virus (EBV) in Guangdong province detected in this study is wt-Xho Ⅰ, while Xho-Ⅰ-loss is mainly found in NPC tissues. Therefore, we think that the loss of Xho Ⅰ restriction site of N-terminal region of Epstein-Barr virus LMP1 gene and its?