青岛地区不同宫颈病变组织中HPV16 E2基因突变分析

Analysis of the mutations of HPV16 E2 gene amony patients with different grades of cervical lesions in Qingdao

目的:探讨山东省青岛地区不同宫颈病变组织中人乳头瘤病毒16型(HPV16)E2基因序列多态性,以及序列变异与不同宫颈病变的关系。方法:提取257例宫颈脱落细胞及宫颈癌组织DNA,PCR进行HPV分型,其中HPV16阳性标本扩增E2基因,测定PCR产物序列。通过DNAStar生物软件进行核苷酸和氨基酸序列分析。结果:共检测出11个碱基突变位点,其中10个导致氨基酸的改变,突变热点为nt3410(63.2%),nt3159(55.9%),nt3249(55.9%)。统计学分析显示,各个位点的突变与不同级别宫颈病变之间均无相关性(P>0.05)。宫颈癌标本中基因整合率为[65%(14/40)],远高于CINⅢ的整合率[14%(6/43)](P<0.001)。结论:青岛地区HPV16 E2基因存在多个位点的突变,且发现G2828A,T3274G,T3384C,T3524C为青岛地区特异性突变,这些位点的突变可能与高度上皮内瘤变和浸润性宫颈癌的发生和发展有关。

Objectives：To investigate the specific nucleotide variations in the human papillomavirus（HPV） 16 E2 DNA sequence in Qingdao of Shan dong Province.The association between the sequence variant and different cervical lesions was also assessed.Methods：The HPV16 E2 gene was amplified by PCR from fresh-frozen tissue as well as cervical-scraping cell lysates and sequenced it and then were compared with the standard strain.Results：A total of 11 nucleotide mutations were identified,while 10 of which resulted in amino acid changes.The hot spots of mutation were nt,3410（63.2%）,3159（55.9%）,3249（55.9%）.The analysis revealed that there was no relation between the E2 mutation and degree of dysplasia（P0.05）.The integration ratio was 65%（14 of 40） in the cervical cancer specimens,significantly more frequent than in CINⅢ lesions（6 of 43）（P0.001）.Conclusion：In this study,there are several E2 sequence variations in HPV16 isolates in qingdao of china,G2828A,T3274G,T3384C,T3524C are found for the first time.These variants may be associated with the developing of cervical cancer.