吉林省宫颈病变患者人乳头瘤病毒16型E6、E7基因变异分析

Analysis of gene variation of human papillomavirus type 16 E6 and E7 in patients with cervical lesions in Jilin Province

目的通过调查吉林省人乳头瘤病毒16型(HPV16)的E6、E7基因变异,评估HPV16基因变异与吉林省地区宫颈癌前病变及宫颈癌发生的相关风险。方法收集129例HPV16感染的宫颈癌及癌前病变患者的宫颈脱落细胞,提取细胞DNA,以特异性引物用聚合酶链反应法(PCR)扩增E6-E7基因序列,再将E6-E7基因序列与标准的HPV16型基因序列比对,寻找E6-E7基因的突变位点。结果共发现8个E6突变位点,其中错义突变6个,无义突变位点2个,突变发生率较高的突变位点为:T178G(59/129,45.74%),A131C(10/129,7.75%),T350G(6/129,4.65%),T178A(6/129,4.65%)。共发现11个E7基因突变位点,其中错义突变位点6个,无义突变位点5个,突变率较高的位点为A647G(57/129,44.19%),T846C(55/129,42.64%),G666A(34/129,26.36%),T843C(20/129,15.50%)。E6、E7常见的突变位点在宫颈癌组和癌前病变组分布均差异无统计学意义。本研究中仅发现了亚洲变异型和欧洲变异型。经统计学分析发现,吉林省HPV16原型、As、Eur在宫颈癌和癌前病变中的分布差异有统计学意义(P=0.02),其中以亚洲变异型为主。结论本研究中吉林省仅发现HPV16的As、Eur2种变异型,且以As变异型为主;在吉林省HPV16 E6、E7常见的变异位点与宫颈病变的进展无关。

Objective To evaluate the relationship between human papillomavirus 16(HPV16)type gene variation and the risk of cervical precancerous lesions and cervical cancer in Jilin Province by investigating the E6 and E7 gene mutations of HPV16 in Julin Province.Methods Cervical exfoliated cells were collected from 129 patients with cervical cancer and precancerous lesions infected with HPV16,DNA was extracted from the cells,and E6-E7 gene sequence was amplified by polymerase chain reaction(PCR)with specific primers,then the E6-E7 gene sequence was compared with the standard HPV16 gene sequence to find the mutation sites of E6-E7 gene.Results A total of 8 mutation sites were found in E6,including 6 missense mutation sites and 2 nonsense mutation sites.The mutation sites with higher mutation incidence were T178G(59/129,45.74%),A131C(10/129,7.75%),T350G(6/129,4.65%),and T178A(6/129,4.65%).A total of 11 gene mutation sites were found in E7,including 6 missense mutation sites and 5 nonsense mutation sites.The mutation sites with higher mutation incidence were A647G(57/129,44.19%),T846C(55/129,42.64%),G666A(34/129,26.36%),and T843C(20/129,15.50%).There was no statiscally significant difference in the distribution of common mutation sites of E6 and E7 between cervical cancer group and precancerous lesion group.Only Asian variants and European variants were found in this study.Statistical analysis showed that there was statiscally significant difference in the distribution of HPV16 prototype,As and Eur in cervical cancer and precancerous lesions in Jilin Province(P-0.02),with the Asian variant being the main type.Conclusion In this study,only As and Eur variants of HPV16 are found in Jilin Province,with As variants being the main type.The common variation sites of HPV16 E6 and E7 in Jilin Province are not associated with the progression of cervical lesions.