青岛地区子宫颈病变组织中人乳头瘤病毒16型L1基因多态性分析

Analysis of HPV16 L1gene variants of different cervical lesions in Qingdao

目的分析青岛地区不同宫颈病变组织中人乳头瘤病毒(HPV)16型L1基因序列的多态性,探讨其序列变异与宫颈癌的相关性。方法提取2008年6月至2010年5月青岛大学医学院附属医院妇产科61例宫颈癌组织和181例宫颈脱落细胞DNA,采用PCR技术筛选HPV16阳性标本,进而对其L1基因全长进行扩增、测序,应用DNASTAR软件进行序列比对,分析核苷酸和氨基酸的变异。结果慢性宫颈炎组、宫颈上皮内瘤变(CIN)组和宫颈癌组HPV16的感染率分别为30.0%(12/40)、41.1%(58/141)和65.6%(40/61),3组比较,差异有统计学意义(P<0.05)。共有96例成功扩增出HPV16L1基因,慢性宫颈炎、CIN和宫颈癌组扩增率分别为25.0%(10/40)、37.6%(53/141)和54.1%(33/61)。HPV16型L1基因共发现13处变异,宫颈癌组织均发生C6240G、A6432G、6902insATC、6954delGAT及G7061A(无义)5处突变,另一突变热点为A6178C,但其在慢性宫颈炎、CIN、宫颈癌组的频率分布比较,差异无统计学意义(P>0.05)。结论青岛地区HPV16 L1基因的多态性具有地域性,可能导致病毒地方株的免疫原性和抗原性改变,但与宫颈癌的发生发展无明显关系。

Objective To analyze sequence variants of the HPV16late gene L1derived from Qingdao women with different cervical lesions,and to detect the relationship between the sequence variant and the cervical cancer.Methods DNA was extracted from 61cervical cancer biopsies and 181cervical-scraping cell lysate.The full length of L1gene was amplified and sequenced,and the DNASTAR software was used to analyze the variants of nucleotide and acid amino.Results The positive rate of HPV16was 30.0% （12/40）,41.1% （58/141）and 65.6% （40/61）in cervicitis,CIN and cervical cancer.The distribution showed statistical difference（P〈0.05）.A total of 96samples were amplified successfully in all samples.The rate were 25.0% （10/40）,37.6% （53/141）and 54.1% （33/61）in cervicitis,CIN and cervical cancer.A total of 13variants were found in the full length of L1gene.Five mutations,including C6240G,A6432G, 6902insATC,6954delGAT and G7061A （nonsense）,were found in all of the positive samples.Mutation of A6178Cwas another hot spot,whose distribution showed no significant difference among cervicitis,CIN and cervical cancer（P〉0.05）.Conclusion There are unique feature of the HPV16L1gene variation,which may cause certain changes concerning immunogenicity and antigenicity of the virus,nevertheless has no apparent connection to the development of cervical lesions.