Zhang Fan, Ding Xiao-hua, Cai Hong-bing, Wang Hua, Chen Chang-chun, Zhang Shuai
Department of Gynecologic Oncology, Zhongnan Hospital, Wuhan University, China.
Zhonghua Zhong Liu Za Zhi. 2010 Feb;32(2):107-10.
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.
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 III 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 HPV16 E6 and E7 specific primers.
The positive rate of E6 gene in cervicitis, CINI and CINII, 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 case 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 cervicitis and CIN I approximately III samples was 25.0% and 31.8%. E7 mutations were found in Nt 647 in 21 cervical samples from 30 cervical cancer samples, resulting in amino acid change of Asn to Ser. The mutation rate was 70.0%. The mutation rate of E6 647 in cervicitis and CINI approximately III samples was 35.0% and 40.9%, respectively.
The positive rate of E6 and E7 increase gradually from cervicitis, CINI and CINII, CINIII 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 HPV16 variants prevalent in this area are different from the European and African variants.
本研究旨在分析人乳头瘤病毒16型（HPV16）转化基因E6、E7和E5在宫颈病变不同阶段的相对分布及基因变异情况。
从200例宫颈病变患者的组织样本中提取DNA，其中包括124例宫颈癌患者、17例宫颈上皮内瘤变（CIN）I级和II级患者、23例CIN III级患者以及36例宫颈炎患者。随后对HPV16 E6、E7和E5基因进行扩增，并使用HPV16 E6和E7特异性引物对部分E6和E7聚合酶链反应（PCR）产物进行测序。
宫颈炎、CIN I级和II级、CIN III级以及宫颈癌中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%。在80例宫颈癌样本中的47例样本中发现HPV16 E6基因第178位核苷酸（Nt）发生突变，导致氨基酸由天冬氨酸（Asp）变为谷氨酸（Glu），突变率为58.8%。此外，宫颈炎和CIN I级至III级样本中E6 178的突变率分别为25.0%和31.8%。在30例宫颈癌样本中的21例宫颈样本中发现E7基因第647位Nt发生突变，导致氨基酸由天冬酰胺（Asn）变为丝氨酸（Ser），突变率为70.0%。宫颈炎和CIN I级至III级样本中E6 647的突变率分别为35.0%和40.9%。
从宫颈炎、CIN I级和II级、CIN III级到宫颈癌，E6和E7的阳性率逐渐升高。在宫颈组织样本中，E5的阳性率相对低于E6和E7基因。这些结果表明，E6和E7基因与宫颈癌的进展高度相关，而E5基因在宫颈癌的发生发展过程中缺失。在中国湖北省的宫颈癌样本中发现HPV16 E6和E7基因在E6 178、E7 647位点存在高频突变。这些结果证实该地区流行的HPV16变异株与欧洲和非洲的变异株不同。
