辽宁省地区人乳头瘤病毒33型基因多态性分析  

Variability in E6 and E7 of Human Papillomavirus Type 33 in Liaoning Province,China

作　　者：于淼[1] 吴思[1] 王爽[1] 崔畅婉 路一平 孙峥嵘[1] YU Miao;WU Si;WANG Shuang;CUI Changwan;LU Yiping;SUN Zhengrong(Biobank of Shengjing Hospital Affiliated to China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院生物样本库,沈阳110004

出　　处：《病毒学报》2022年第2期354-362,共9页Chinese Journal of Virology

基　　金：辽宁省重点研发计划(项目号:2018225009),题目:人乳头瘤病毒16型E7蛋白通过选择性调控干扰素刺激基因群ISGs表达抵抗β干扰素抗病毒作用。

摘　　要：研究人乳头瘤病毒(Human papillomavirus,HPV)33型E6和E7基因在辽宁省地区的基因多态性分布情况,增加对HPV33基因多态性地理分布情况的了解,为HPV33的诊断,靶向药和疫苗的研发提供理论依据。使用巢式PCR扩增方法对从已确诊HPV33阳性患者宫颈细胞提取的DNA样本进行E6和E7基因的扩增并测序,随后应用MegaX软件将所得测序结果与参考序列比对确定突变位点并进行系统发育树的构建。应用PAML4.9软件中的Codeml程序找出突变中的正向选择位点。分别应用ABC Pred server,ProPred-I server和ProPred server寻找理想的B细胞免疫表位,人类白细胞抗原(Human leukocyte antigen,HLA)I类结合表位和人类白细胞抗原(HLA)II类结合表位。共得到HPV33E6和E7序列各136个。与HPV33参考序列(M12732.1)进行比对后,E6基因中观测到17个单核苷酸突变,同义突变7个,非同义突变10个。E7基因中观测到9个单核苷酸突变,其中同义突变3个,非同义突变6个。系统发育分析显示HPV33E6和E7 A谱系在中国东北地区最为流行,其次是B谱系,本研究中并未检测到C谱系。在全部E6和E7序列中仅找到K93N一个阳性选择位点位于E6基因。在HPV33E6中预测得到8个MHCI类表位,6个MHCII类表位和1个B细胞表位。HPV33E7中预测到5个MHCI类表位,6个MHCII类表位和3个B细胞表位。研究中检测到的绝大部分非同义突变存在于预测表位中。在辽宁省地区,HPV33E6和E7基因呈基因多态性分布,且A谱系为最常见谱系。研究所得结果可为HPV33检测探针、靶向药物和疫苗的研究和临床应用提供理论基础。To study the polymorphisms of E6 and E7 in human papilloma virus(HPV)33 genes in Liaoning Province,China;increase understanding of the geographic distribution of polymorphisms of the HPV33 gene;provide a theoretical basis for the development of the diagnosis,targeted drugs and vaccines of HPV33. DNA samples extracted from the cervical cells of HPV33-positive patients were amplified using nested polymerase chain reaction(PCR)amplification and then sequenced. After that,MegaX was used to compare the sequences with the reference sequence to determine the mutation site and construct phylogenetic trees. We used the“Codeml”program in PAML4.9 to find the positive selection site in the mutation. Several servers(ABC Pred,ProPred-I,ProPred)were employed to find the“ideal”B-cell immune epitope,as well as human leukocyte antigen(HLA)class-I and class-II binding epitopes,respectively. A total of 136 HPV33 E6 and E7 sequences were obtained. Seventeen single-nucleotide mutations were observed in E6:seven were synonymous mutations and 10 were non-synonymous mutations. Nine single-nucleotide mutations were observed in E7:three were synonymous mutations and six were non-synonymous mutations. Phylogenetic analysis showed that lineage A of E6 and E7 of HPV33 was most prevalent in Northeast China,followed by lineage B. No sequence belonging to lineage C was not detected. Among all E6 and E7 sequences,only one positive selection site of K93 N was found in E6. In E6 of HPV33,eight MHCI epitopes,six MHCII epitopes and one B-cell epitope were predicted. In E7 of HPV33,five MHC class-I epitopes,six MHC class-II epitopes and three B-cell epitopes were predicted. The vast majority of non-synonymous mutations detected in our study were presented in the predicted epitopes. In Liaoning Province,E6 and E7 of HPV33 were polymorphic,and lineage A was the most common lineage. These results provide a theoretical basis for the research and clinical application for detection probes,targeted drugs and vaccines for HPV33.

关 键 词：HPV33 E6基因 E7基因 基因多态性 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]