HPV16 E2基因各区域缺失与宫颈病变相关性的研究

目的:检测湖北地区HPV16阳性宫颈标本E2基因碳端(C)、铰链区(H)、氮端(N)的缺失状态,探讨其与CIN及宫颈癌的关系。方法:将122例宫颈标本按病变程度分为宫颈癌组、CINⅡ~Ⅲ组、对照组。应用多重聚合酶链反应(PCR)将各组HPV16感染标本E2基因的C、H、N分别与E6基因同时扩增,检测E2基因各区域的缺失状态,并应用Scion Image4.0软件半定量分析E2、E6基因条带灰度值,判断HPV DNA的整合状态。结果:宫颈癌组E2基因C、H、N缺失率分别是22.5%,50%,77.5%;CIN组为30.77%,69.23%,92.31%;对照组为9.09%,27.27%,27.27%。E2基因C、H缺失率在3组的任两组间比较无统计学差异(P0.05),而E2基因N端缺失率在宫颈癌组与对照组之间及CIN组与对照组之间差异有统计学意义(P均0.01)。结论:HPV16病毒E2基因N端缺失状态与CIN及宫颈癌有内在关系。在检测HPV感染同时检测E2基因N端缺失状态,对间接判断预后及指导后续治疗有重要意义。

Different deletion states of HPV16 E2 gene in C-terminal, hinge region and N-terminal in cervical lesions

Objective:To detect the different deletion state of HPV E2 gene in C terminal(C),hinge region(H),N terminal(N)in HPV16-positive cervical samples,and investigate the association between the deletion state and CIN or cervical cancer.Methods:According to different levels of disease,122 cervical samples were divided into three groups:cervical cancer group,CINⅡ~Ⅲ group and control group.The specimens which infected by HPV16 were amplified by multiple polymerase chain reaction(PCR),using three overlapping fragments amplified with HPV E6 gene to detect the different miss rate of different position of E2 gene in different groups.Scion Image 4.0 electrophoresis image analysis soft was used to calculate the area of electrophoresis product of E2 gene and E6 gene to analysis the integration state of HPV16.Results:In cervical cancer group,the miss rate of C,H,N are 22.5%,50% and 77.5%;in CIN group are 30.77%,69.23%,92.31% and in control group are 9.09%,27.27%,27.27%.The deletion rate of C or H of HPV16 E2 gene have no significant difference betweens groups(P>0.05),while the deletion rate of N-terminal of HPV16 E2 gene have statistical difference between the cervical cancer group and control group,also between the CIN group and the control,(P<0.01,P<0.01).Conclusion:The N terminal deletion status of HPV16 E2 gene with CIN and cervical cancer are intrinsically linked.We can judge the prognosis and decide the follow-up treatment of patients by detecting HPV and exist state of HPV16 E2 in N-terminal.