人乳头瘤病毒16型转化基因在不同阶段宫颈上皮病变组织中的分布及基因变异特点  

Analysis of the relative distribution and gene variation of HPV16 transforming gene at different stages of cervical lesions

作　　者：张帆[1] 丁晓华[2] 蔡红兵[1] 王华[1] 陈长春[3] 张帅[1] 

机构地区：[1]武汉大学中南医院妇瘤科肿瘤生物学行为湖北省重点实验室,430071 [2]武汉大学医学院医学病毒研究所 [3]湖北省肿瘤医院CT室

出　　处：《中华肿瘤杂志》2010年第2期107-110,共4页Chinese Journal of Oncology

基　　金：湖北省卫生厅科研基金（JX3A17）

摘　　要：目的研究人乳头瘤病毒（HPV）16E6、E7和E5基因在湖北地区不同阶段宫颈上皮病变患者组织中的分布以及E6、E7基因的变异特点。方法从124例宫颈癌、17例宫颈上皮内瘤变（CIN）Ⅰ＋CINⅡ级、23例CINDⅠ级和36例慢性宫颈炎患者活检或手术切除标本中提取组织DNA，用HPV16E6、E7和E5特异性引物进行PCR扩增，对部分扩增的E6和E7产物片段进行测序分析。结果在宫颈炎、CINⅠ＋CINⅡ级、CINⅢ级和宫颈癌组织中，E6基因的阳性率分别为25．0％、29．4％、60．9％和76．6％；E7基因的阳性率分别为16．7％、41．2％、43．5％和61．3％；E5基因的阳性率分别为5．6％、5．9％、30．4％和40．3％。E6、E7和E5基因在不同阶段宫颈上皮病变组织中的阳性率差异均有统计学意义（均P〈0．01）。在80例宫颈癌测序组织中，有47例发生E6基因178位点的T—C突变，突变率为58．8％，相应氨基酸由天冬氨酸（Asp）改变为谷氨酸（Glu）；而在20例宫颈炎和22例CINⅠ～Ⅲ级测序组织中，E6基因178位点的突变率分别为25．0％和31．8％。在30例宫颈癌测序组织中，有21例发生E7基因647位点的A—G突变，突变率为70．0％，相应氨基酸由天冬酰胺（Asn）改变为丝氨酸（Ser）；而在20例宫颈炎和22例CINⅠ～Ⅲ级测序组织中，E7基因647位点的突变率分别为35．0％和40．9％。结论HPV16E6、E7和E5基因与宫颈癌的发生和发展有高度的相关性，但E5基因在不同阶段宫颈上皮病变中可能存在不同程度的缺失。中国湖北地区流行的HPV16病毒株可能为HPV16亚洲型变异株。Objective The aim of this study was to analyze the relative distribution and gene variation of HPV16 transforming gene E6, E7 and E5 at different stages of cervical lesions. Methods DNA was extracted from tissue samples of 200 patients with cervical lesions, including 124 cervical cancer, 17 CIN grade Ⅰ and Ⅱ, 23 CIN grade m and 36 cervicitis. Then HPV16 E6, E7 and E5 genes were amplified, and part of the E6 and E7 PCR products were sequenced using the HPVl6 E6 and E7 specific primers. Results The positive rate of E6 gene in ccrvicitis, CIN Ⅰ and CIN Ⅱ , CINⅢ and cervical cancer was 25.0% ,29.4% ,60.9% and 76.6% , respectively. The positive rate of E7 gene was 16.7% ,41.2% , 43.5% and 61.3% , respectively. The positive rate of E5 gene was 5.6% ,5.9% ,30.4% and 40.3% , respectively. HPV16 E6 gene mutations in Nt 178 were found in 47 ease from 80 cervical cancer samples, resulting in amino acid change of Asp to Glu. The mutation rate was 58.8%. Otherwise the mutation rate of E6 178 in cervieitis and CIN Ⅰ -Ⅲ samples was 25.0% and 31.8%. E7 mutations were found in Nt 647 in 21 cervical samples from 30 cervical cancer samples, resuhing in amino acid change of Ash to Ser. The mutation rate was 70.0%. The mutation rate of E6 647 in cervieitis and CIN Ⅰ-Ⅲ samples was 35.0% and 40.9%, respectively. Conclusion The positive rate of E6 and E7 increase gradually from cervicitis, CIN Ⅰ and CINⅡ, CIN Ⅲ to cervical cancer. The rate of E5 is relatively lower than that of E6 and E7 gene in cervical tissue samples. These results show that E6 and E7 gene are highly associated with the progress of cervical cancer and E5 genes are lost in the development of cervical cancer. High frequency mutations of HPV16 E6 and E7 gene in E6 178, E7 647 have been found in cervical cancer samples in Hubei province, China. These results approved that the HPVI6 variants prevalent in this area are different from the European and African variants.

关 键 词：宫颈疾病 人乳头瘤病毒16 序列分析 突变 

分 类 号：R737[医药卫生—肿瘤]