新疆维、汉族宫颈鳞癌病变中HPV16 E6基因变异分析

Analysis of HPV16-E6 gene mutation in Xinjiang Uygur and Han women with cervical squamous carcinoma

目的分析新疆地区维、汉族妇女宫颈鳞癌组织中HPV16型E6基因的变异情况。方法从新疆地区5例维吾尔族妇女和5例汉族妇女宫颈活检组织中提取病毒基因组DNA,经反向膜杂交检测确定为HPV16单一感染。采用PCR技术扩增HPV16E6基因片段,克隆到pUC18-T载体中进行DNA测序。结果成功克隆了HPV16E6基因,DNA测序结果表明,与德国标准株相比,10例宫颈鳞癌组织中共检出12种E6基因突变类型,包括107(T→C)、133(A→C)、168(C→G)、178(T→G)、310(T→C)、321(T→C)、341(T→C)、350(T→G)、375(A→G)、410(T→A)、499(G→T)和548(A→G),其中178(T→G)和350(T→G)的突变频率较高,均为30%(3/10)。汉族和维吾尔族标本中共有的突变包括178(T→G)和350(T→G),仅存在于汉族标本的突变为107(T→C)、133(A→C)、168(C→G)、310(T→C)和499(G→T),仅发生在维吾尔族标本的突变为321(T→C)、341(T→C)、375(A→G)、410(T→A)和548(A→G)。结论与德国标准株比较,新疆维吾尔族、汉族宫颈癌患者中HPV16E6基因存在新的变异,其中178(T→G)和350(T→G)型基因变异为热点突变;新疆地区宫颈癌的高发可能与HPV16E6基因突变有关。

Objective The present study investigates the HPV16-E6 gene mutation in Xinjiang Uygur and Han women with cervical squamous carcinoma.Methods The viral genome DNA was extracted from the cervical biopsies of five Uygur and five Han patients from Xinjiang,and the disease was determined to be an HPV16 simplex infection through the reverse membrane hybridization test.PCR was used to amplify the HPV16-E6 genetic fragment that is to be cloned into a pUC18-T carrier for DNA sequencing.Results The HPV16-E6 gene was cloned successfully.The DNA sequencing results showed that,compared with the DNA sequence of the German strain of the HPV16-E6 gene,12 mutation types were detected in the 10 specimens.These types were 107（T→C）,133（A→C）,168（C→G）,178（T→G）,310（T→C）,321（T→C）,341（T→C）,350（T→G）,375（A→G）,410（T→A）,499（G→T）,and 548（A→G）.Among them,178（T→G） and 350（T→G） had higher mutation frequencies,which were 30%（3/10） and 30%（3/10）,respectively.Furthermore,data analysis showed that 178（T→G） and 350（T→G） existed in both Uygur and Han women.However,107（T→C）,133（A→C）,168（C→G）,310（T→C）,and 499（G→T） were detected only in Han women,whereas 321（T→C）,341（T→C）,375（A→G）,410（T→A）,and 548（A→G） were identified only in Uygur women.Conclusions New HPV16-E6 gene variations were identified in Xinjiang Uygur and Han women with cervical cancer by comparing their genes with the German standard HPV16-E6 gene.Moreover,178（T→G） and 350（T→G） were found to be the hot spots of the HPV16-E6 gene mutation in these women.Therefore,the high incidence of cervical cancer in Xinjiang Uygur and Han women may be associated with the HPV16-E6 gene mutation.