成渝两地宫颈癌患者中人乳头瘤病毒16亚型L1、E6和E7基因的分离克隆和序列分析  

Isolating,Cloning,and Sequence-analyzing of HPV16 L1、E6 and E7 Gene from Cervical Carcinoma Biopsis in Southwestern China

作　　者：陈良[1] 丁显平[1] 朱一剑[1] 李付广[1] 

机构地区：[1]四川大学生命科学学院遗传医学研究所,生物资源与生态环境教育部重点实验室,四川成都610064

出　　处：《国际检验医学杂志》2010年第3期217-219,共3页International Journal of Laboratory Medicine

摘　　要：目的了解成渝地区HPV16亚型基因的一级结构特点,为成渝地区宫颈癌的防治及HPV疫苗的研制提供理论基础。方法收集宫颈癌组织标本,针对HPV16亚型分别设计L1、E6和E7基因特异性引物,用PCR方法从宫颈癌组织中获得各基因的全长序列,并构建克隆载体pGEM-T,进行序列分析,同时分别以L1、E6和E7基因序列进行进化分析。结果在102例宫颈癌标本中,99例为HPV阳性,感染率为97.1%,其中32例(32.3%)为HPV16单重感染,所占比例最高。通过与标准株比较,发现成渝两地HPV16亚型E6、E7和L1基因都存在一定程度的变异,变异率分别为90.6%、78.1%和25.0%。结论与标准序列比较,成渝地区HPV16亚型L1、E6和E7基因存在一定范围的变异,提示我们,在预防治疗成渝地区妇女宫颈癌病人以及研制开发针对西南地区的宫颈癌疫苗时,需要注意这些位点的变异,特别是一些引起氨基酸变异的位点。Objective To know the primary structure character of HPV 16 isoforms in CHENG-YU district, providing the foundation for preventing and curing the cervical carcinoma. Methods E6、E7 and L1 gene were amplified from human cervical carcinoma by PCR. The products of PCR were ligated into pGEM-T vector and used to transform E. coli DHSα. At last, the sequence of the insert was determined by an automated DNA sequencer. Results Of the 102 cervical carcinoma specimens, 97.1% （99/102） were positive for HPV infection, including 32.3% （32/99） HPV16 single infection. Compared with the prototype, a certain variation was found in HPV isoforms of E6, E7, L1, the variation ratio was 90.6%, 78.1%, and 25.0% respectively. Conclusion Compared with the prototype, the nucleotide sequence of the HPV16 E6. E7 and L1 gene from cervical carcinoma biopsis in CHENG-YU district shows a certain point mutations. We should pay attention to this mutations in preventing and curing cervical carcinoma in CHENG-YU district.

关 键 词：人乳头瘤病毒16 宫颈肿瘤 序列分析 

分 类 号：R737.33[医药卫生—肿瘤]