宫颈癌组织HPV16 URR及E7基因突变和序列多态性分析_医学论文.doc免费

宫颈癌组织HPV16 URR及E7基因突变和序列多态性分析_医学论文 宫颈癌组织HPV16 URR及E7基因突变和序列多态性分析_医学论文 作者：赵雪珍 王言奎 彭丽娜 张红 【摘要】 目的 分析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E7开放读码框(OFR)基因突变和基因序列多态性。方法 收集山东省青岛地区妇女宫颈癌标本共111例，提取DNA作为模板，应用通用引物和型特异引物经聚合酶链反应(PCR)筛选HPV16阳性标本，PCR结合测序检测HPV16阳性标本URR和E7基因全序列，经DNAStar 100软件和在线BLAST分析其变异情况。结果 宫颈癌组织中HPV和HPV16感染率分别为99.10%(110/111)和73.63%(81/110)核苷酸水平上HPV16 URR形成24种变异模式，同源性在98.02%～99.26%之间，nt7518 G→A和nt7861 A缺失变异率均为100%(26/26)。HPV16 E7形成12种变异模式，同源性在99.23%～100%之间nt647 A→G突变率58.54%(24/41)，氨基酸由Asn→Ser(N29S)、nt666 G→A突变率24.39%(10/41)，为同义突变。结论 山东省青岛地区妇女宫颈癌组织中HPV16 URR和E7基因均存在变异HPV16 URR突变热点为nt7518和nt7861，HPV16 E7突变热点为nt647，且二者尚存在未见报道的变异位点。 【关键词】 宫颈肿瘤人乳头瘤病毒16调控序列,核酸聚合酶链反应 [ABSTRACT] Objective To analyze the mutation and polymorphism of HPV16 URR and E7 gene in cervical cancer samples in Qingdao of Shandong province. Methods A total of 111 cancer samples were collected in Qingdao. The DNA was extracted and used as a template. The detection of HPV and the typing of HPV16 carried out using consensus and typespecific primers. The amplifications of HPV16 URR and E7 gene complete sequences were performed by PCR with the HPV16 positive ones. The variation information was analyzed by DNAStar 100 and BLAST online. Results The positive rate of HPV and HPV16 was 99.10% (110/111) and 73.63% (81/110), respectively. HPV16 URR fragments formed 24 patterns at nucleotide level, their homology was 98.02% to 99.26%. nt7518 G→A and nt7861 A absence were 100% (26/26) HPV16 E7 fragments formed 12 patterns mutation at amino acid level, their homology was 99.23% to 100% HPV16 E7 nt647 A→G was 58.54% (24/41), resulting in amino acid Ser substituted for Asn. nt666 G→A was 24.39% (10/41), without variation of amino acid. Conclusion There are variations in both HPV16 URR and E7 genes at nucleotide level in Qingdao. The mutational hot spots of HPV16 URR are nt7518 and nt7861 and HPV16 E7 is nt647, both of them still have mutational spots that have not yet been reported. 　　[KEY WORDS] uterine c