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目的:分析湖北地区宫颈癌组织HPV16E7和E5基因序列变异情况。方法:对87例宫颈鳞癌组织分别采用多重HPV16E7引物进行巢式PCR扩增和HPV16E5特异引物进行PCR扩增,并选择阳性扩增的基因片段DNA进行纯化、测序,检测基因变异。结果:测序成功的20例宫颈癌组织DNA中,HPV16E7基因的5个突变位点为:T846C、A647G、T732C、T789C、T795G,突变频率分别为85%(17/20)、70%(14/20)、15%(3/20),10%(2/20)、5%(1/20);HPV16E5基因的4个突变位点包括:A3979C、A4042G、G3868A、C3991T,突变频率分别为:80%(16/20)、90%(18/20)、5%(1/20)、5%(1/20)。结论:湖北地区宫颈癌的宫颈组织中HPV16E7热点突变为A647G和T846C,HPV16E5的热点突变为A3979C和A4042G,本研究为湖北地区宫颈癌流行病学的研究打下基础。
Objective: To analyze the variation of HPV16E7 and E5 gene sequences in cervical cancer in Hubei province. Methods: 87 cases of cervical squamous cell carcinoma were amplified by PCR with multiple HPV16E7 primers and HPV16E5-specific primers respectively. The gene fragments were selected and amplified by sequencing. Results: The mutations of HPV16E7 gene in the DNA of 20 cervical cancer tissues sequenced successfully were 85% (17/20) and 70% (14%) of T846C, A647G, T732C, T795C and T795G, 20, 15% (3/20), 10% (2/20) and 5% (1/20) respectively. The four mutation sites of HPV16E5 gene include A3979C, A4042G, G3868A and C3991T, 80% (16/20), 90% (18/20), 5% (1/20), 5% (1/20). Conclusion: The hot spots of HPV16E7 mutation are A647G and T846C in cervical tissues of Hubei province. The hot spot mutations of HPV16E5 are A3979C and A4042G. This study lays the foundation for the epidemiological study of cervical cancer in Hubei province.