人乳头瘤病毒18型E6/E7基因在宫颈脱落细胞中的表达研究  

Genetic variability of Human Papillomavirus 18 E6/E7 gene and its expression in cervical exfoliated cell

作　　者：汪萍 宛传丹 崔燕红 赵一琳 丘佳明[3] WANG Ping;WAN Chuan-dan;CUI Yan-hong;ZHAO Yi-lin;QIU Jia-min(Basic Medical department of Wannan Medical College, Anhui, Wuhu 241002, China)

机构地区：[1]皖南医学院基础医学院医学生物学教研室,安徽芜湖241002 [2]常熟市医学检验所分子生物学实验室,江苏常熟215500 [3]常熟市第二人民医院病理科,江苏常熟215500

出　　处：《现代预防医学》2018年第10期1908-1910,1920,共4页Modern Preventive Medicine

基　　金：安徽高校省级优秀青年人才基金项目(2012SQRL123)

摘　　要：目的研究人乳头瘤病毒HPV 18型早期基因E6/E7在不同宫颈上皮病变患者脱落细胞中mRNA表达分布与基因突变特征。方法收集单纯感染HPV 18型不同阶段宫颈上皮病变组织脱落细胞并提取总DNA。特异性引物针对20例宫颈癌患者E6、E7基因进行PCR扩增检测,所得产物进行Sanger测序分析。以支链DNA(b-DNA)技术检测HPV18型E6/E7 mRNA阳性率,与临床病理CIN结果进行比较分析。结果基因测序发现20例发生E6基因突变,199位点突变率最高,达55.0%(11/20);再次是201位点突变率45.0%(9/20)。15例发生E7基因突变,其中125位点突变率60.0%(9/15),199位点突变率33.3%(5/15)。在不同宫颈上皮病变患者脱落细胞标本中,E6/E7 mRNA阳性率呈现差异性(P<0.01)。E6/E7 mRNA阳性率在慢性宫颈炎、CIN Ⅰ+Ⅱ级、CIN Ⅲ级和宫颈癌组织中阳性率分别为23.4%、35.7%、81.1%和95.1%。E6/E7 mRNA阳性率与临床CIN分级诊断呈正向线性相关。结论 HPV 18型基因突变较为突出。HPV18型早期蛋白E6、E7表达量的变化与宫颈组织病变程度高度相关。Objective To analyze the expression and variability of HPV 18 transforming early gene E6, E7 in cervical exfoliated cell at different stages. Methods Different stages of cervical exfoliated cells were collected from simple HPV 18 virus infected patients and its DNA was extracted. E6, E7 genes were amplified, and their products in cervical cancer were sequenced using specific primers in 20 cases of cervical cancer. Branded DNA（b-DNA） technology was used to evaluate the relationship of HPV 18 E6/E7 mRNA positive rate and the clinical diagnosis of CIN. Results E6 gene mutation was found in 20 cases. The highest mutation rate was 55%（11/20） in the 199 locus, and the mutation rate was 45%（9/20） in the 201 loci.Mutation of E7 gene occurred in 15 cases, the mutation rate of 125 was 60%（9/15）, and the mutation rate of 199 was 33.3%（5/15）. The expressions of E6/E7 mRNA of HPV 18 demonstrated difference at different stages of cervical exfoliated cells（P0.05）. The positive rate of E6/E7 mRNA was 23.4%, 35.7%, 81.1% and 95.1% in cervicitis, CIN Ⅰ and CIN Ⅱ, CIN Ⅲ and cervical cancer respectively. The positive rate of E6/E7 mRNA was positively correlated with clinical CIN classification.Conclusion HPV 18 gene mutation proceeds at our studied regions. Early gene E6, E7 in HPV 18 are highly associated with the progression of cervical cancer.

关 键 词：人乳头次瘤病毒18型 宫颈癌 脱落细胞 早期基因 突变 

分 类 号：R181.3[医药卫生—流行病学]