青岛地区宫颈癌组织中HPV16多态性分析

Analysis of variants of human papillomavirus type 16 in women with cervical cancer in Qingdao

目的探讨山东省青岛地区妇女宫颈癌组织中人乳头瘤病毒16型(HPV16)上游调控区(URR)和E6基因序列多态性,病毒主要分支,以及他们与宫颈癌的相关性。方法从43例HPV16阳性宫颈癌组织中提取DNA,扩增URR及E6基因,进行序列测定。通过DNASTAR生物软件进行核苷酸和氨基酸序列的分析。结果所有标本URR测序结果均发生了G7521A位点突变;26例(60.5%)宫颈癌组织中检测到核苷酸位点C24T,A7730C和G7842A的联合突变,为另一突变热点。E6测序结果发现突变热点为T178G(D25E),突变率为65.1%(28/43);仅1例T350G突变(L83V)。AS型是最多见的病毒突变类型,占65.1%(28/43),其次为野生型(E-P)23.3%(10/43)。结论青岛地区妇女宫颈癌组织中HPV16URR突变热点为G7521A以及C24T,A7730C和G7842A的联合突变。HPV16E6突变热点为T178G,这些突变热点可能与宫颈癌发生发展有密切关系。AS和E-P原型是青岛地区宫颈癌组织中两种主要的HPV16分支。

Objective To investigate the genetic mutations and co-mutations in the E6 and URR genes of HPV16. Methods Fresh-frozen tissues were obtained from 43 cervical cancer patients. We amplified the HPV16 URR and E6 gene by PCR from these tissues and sequenced it. The products of PCR were purified and sequenced,and then were compared with the standard strain.Results The most mutational hot spot of HPV16 URR was m7521.A remarkable finding was the co-mutations including nt 24,nt 7730 and nt 7842. The most frequent sequence variation of E6 was nt 178 （65.1%）. The mutation of L83V was only found in 1 case. In Qingdao area,two major branches of HPV16 were type AS 65.1%（28/43） and proptype 23.3%（9/26）.Conclusions The mutational hot spots of HPV16 URR are nt 7521 and a co-mutation including nt24,nt7830 and nt7842.The hot spot of E6 was nt178. These variants may be associated with the developing cervical cancer. HPV16AS lineage was predominant in Qingdao.