宫颈癌妇女人乳头瘤病毒16E6/E7基因变异分析

Analysis of human papillomavirus 16 E6/E7 gene mutation cervical cancer

目的了解宫颈癌妇女人乳头瘤病毒(HPV)16E6/E7基因变异的情况,为HPV防治提供流行病学数据。方法从大量样本中筛选出HPV16阳性标本180例,其中宫颈癌患者100例,宫颈不典型增生患者50例,宫颈炎症患者30例,对宫颈脱落细胞样本采用PCR方法扩增HPV16 E6/E7基因,然后对反应产物采用DNA测序法获得基因序列,在GeneBank进行经BLAST分析,寻找变异位点。结果宫颈癌患者中E6核苷酸变异率为82.00%(82/100),明显高于宫颈不典型增生患者(=0.000)及宫颈炎患者(=0.000);宫颈癌HPV16E6最频繁的序列变异为T178G(60/100),其他常见变异为T350G、C335T/A442C、T178G/G39T,年轻宫颈癌组E6核苷酸变异率与非年轻宫颈癌无明显差别,但两者T178G变异率存在明显差异(=0.000)。宫颈癌患者HPV16 E7核苷酸变异率为74.00%(74/100),明显高于宫颈不典型增生患者(=0.000)及宫颈炎患者(=0.000);E7最频繁序列变异为A647G 32.00%(32/100),其他常见的变异为A647G/C749T/T843C/T846C、A647G/C749T/T846C、C749T/C790T、A647G/C749T;年轻宫颈癌组E7核苷酸变异率与非年轻宫颈癌无明显差别,但A647G变异率存在明显差别(=0.03)。结论宫颈癌患者明显存在HPV16E6E7核苷酸变异,HPVE6 T178G、E7 A647G可能与宫颈癌年轻化密切相关,故对该人群的检测并及时尽早采取干预措施。

Objective To investigate of E6/E7 gene variation of human papillomavirus（HPV）16 infection in women with uterine cervical cancer for epidemiological data of HPV prevention.Methods HPV16 positive samples were selected from a large number of samples,and then randomly selected cervical cell samples from100 cases of cervical cancer patients,50 cases of cervical dysplasia patients,30 cases of cervical inflammation and amplified HPV16 E6/E7 gene by PCR,then reaction products obtained gene sequences with DNA sequencing and found the variable sites by BLAST analysis in the GeneBank.Results The rate of E6 nucleotide variation in cervical cancer patients was 82.00%（82/100）,significantly higher than that of cervical dysplasia,cervicitis.The most frequent HPV16 E6sequence variation was T178G（60/100）;other common variation as T350G,C335T/A442C,T178G/G39T,young cervical cancer group E6 nucleotide mutation rate was no significant difference to that of non-young cervical cancer,but the T178G mutation rate significantly different.The HPV16 E7 nucleotide variation rate patients with cervical was 74.00%（74/100）,that was significantly higher than cervical dysplasia,cervicitis;the most frequent sequence variation in the E7 was A647G 32.00%（32/100）.Other common variations for A647G/C749T/T843C/T846C,A647G/C749T/T846C,C749T/C790T,A647G/C749T;E7 nucleotide variation rate of the young cervical cancer group was no significantly higher than non-young cervical cancer,but A647G mutation rate wassignificantly differencent.Conclusions It clearly exists of HPV16 E6 E7 nucleotide variation from cervical cancer patient in Wenzhou City and HPV E6 T178G,the E7 A647G may be closely associated with younger cervical cancer,so it was important to detect the population and take timely measures.