人乳头瘤病毒52型E 6和E 7基因突变与宫颈癌及癌前病变关系的研究  

The study of the association of human papillomavirus 52 E 6 and E 7 mutations,cervical cancer and precancer lesions

作　　者：王本敬[1,2,3] 刘敏娟[1,2,3] 李文静[4] 侯顺玉[2] 王挺[1] 戴建荣[2] WANG Ben -ring;LIU Min -juan;LI Wen -jing;HOU Shun - yu;WANG Ting;DAI Jian - rong(Department Center for Reproduction and Genetics;Department of Gynaecology;Department of Central Laboratory;Department of Clinical Laboratory, Suzhou Hospital Affiliated to Nanfing Medical University, Suzhou Jiangsu 215002, P. R. China)

机构地区：[1]南京医科大学附属苏州医院生殖遗传中心,江苏苏州215002 [2]南京医科大学附属苏州医院妇科,江苏苏州215002 [3]南京医科大学附属苏州医院中心实验室,江苏苏州215002 [4]南京医科大学附属苏州医院检验科,江苏苏州215002

出　　处：《中国计划生育和妇产科》2018年第6期24-30,共7页Chinese Journal of Family Planning & Gynecotokology

基　　金：江苏省妇幼保健科研项目(项目编号:F 201432);苏州市临床医学中心(项目编号:Szzx201505);苏州市"科教兴卫"项目(项目编号:kjxw 2014018;kjxw 2015022;SYS 201429);苏州市"重点病种"临床诊疗技术项目(项目编号:LCZX201410)

摘　　要：目的分析人乳头瘤病毒(human papillomavirus,HPV)52型E 6和E 7基因突变位点分布,评估基因突变与宫颈癌及癌前病变的相关性。方法收集2016年1月至2017年5月经HPV DNA检测为HPV 52单独阳性的DNA样本120例。应用Generunner V 3.01软件设计扩增E 6/E 7区域的1对PCR引物和Sanger测序的4条延伸引物。以HPV 52原型(Gen Bank Accession No.X 74481.1)作为参考序列,应用MEGA 5.05软件分析每个样本的碱基序列和蛋白序列的变异;应用SPSS 17.0进行一般线性相关检验和Fisher's检验或者Pearson Chi-square(χ~2)检验,分析E 6和E 7基因突变与宫颈病变的关系。结果在104例样本中,共发现HPV 52 E 6基因的16个突变位点和E 7基因的17个突变位点。HPV 52 E 6和E 7基因中不同突变位点的突变率差异无统计学意义(χ~2=14.829、12.548,P=0.464、0.766)。在HPV 52 E 6和E 7基因的33种突变中,无突变位点的突变率随着宫颈病变等级恶化有升高或降低的趋势(P>0.05)。HPV 52 E 6基因在CIN 1+样本中的突变频率(14.0%)显著低于正常样本中突变频率(34.4%),(OR=0.31,95%CI=0.11~0.85,χ~2=5.500,P=0.019)。E 7基因的突变频率在CIN 1+样本中的突变频率(16.3%)也低于在正常样本中的突变频率(27.9%),但差异无统计学意义(OR=0.50,95%CI=0.19~1.35,χ~2=1.908,P=0.167)。结论在单独感染HPV 52的条件下,E 6基因突变是阻碍宫颈病变和宫颈癌发生发展的因素。Objective To analyze the distribution of human papillomavirus（HPV） 52 E 6 and E 7 mutations and evaluate the association of the gene mutations,cervical cancer and precancer lesions.Methods 65 samples tested HPV 52 single positive were collected during January 2016 to May 2017.One pair of primers targeting HPV 52 E 6/E 7 region for PCR and 4 primers for Sanger sequencing were designed using Generunner V3.01.Compared to protype HPV 52（Gen Bank Accession No.X74481.1）,the E 6 and E 7 mutations were analyzed using MEGA 5.05.For examining distributions of HPV 52 variations with respect to disease severity,the Linear-by-Linear Association and Fisher＇s exact test or Pearson Chi-square（χ2） were employed.Results In 104（83 %） of 120 samples,16 mutations of E 6 gene and 17 mutations of E 7 gene were found.The prevalence of mutations of E 6 and E 7 genes were not significantly different（χ2= 14.829,P = 0.464 for E s6 gene and χ2= 12.548,P = 0.766 for E 7 gene）.In the 33 mutations of HPV52 E 6 and E 7 genes,the mutation rate of no mutation siteg increased of decreased with the deterioration of cervical lesions（P 0.05.The mutation prevalence of E 6 gene in CIN 1＋（14.0 %） was significantly lower than that in normal samples（34.4 %）,（OR = 0.31,95 % CI = 0.11 ~ 0.85,χ2= 5.500,P = 0.019）.The mutation prevalence of E 7 gene in CIN 1＋（16.3 %） was also lower that in normal samples（27.9 %）,but not achieve significant difference,（OR = 0.50,95 % CI = 0.19 ~ 1.35,χ2= 1.908,P = 0.167）.Conclusion In the condition of HPV 52 single infection,the mutation of E 6 gene is a prevention factor for cervical cancer and precancer lesions.

关 键 词：宫颈癌 基因突变 E6基因 E7基因 人乳头瘤病毒52型 

分 类 号：R737.33[医药卫生—肿瘤]