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目的探讨广东地区宫颈癌组织中HPV16肿瘤相关性抗原E6基因序列的多态性及同源性。方法采用通用引物PCR直接测序法对宫颈癌标本中的HPV分型,从含有HPV16型的标本中采用自行设计的多重引物通过巢式PCR扩增出HPV16E6,经DNA序列测定法检测其基因变异,进而分析其同源性。结果50例宫颈癌组织HPV-DNA的检出率为78%,其中HPV16和HPV18型混合感染18例,单纯HPV16型感染15例。含有HPV16型的标本34例中扩增出HPV16E625例。其中178位核苷酸变异较大,变异率为72%,其相应氨基酸均由天冬氨酸变为谷氨酸。结论HPV16E6DNA序列发生碱基替换的区域主要在氨基端94~241位,羧基端相对保守,未见变异。广东地区宫颈癌组织中HPV16E6的热点突变为Nt178。
Objective To investigate the polymorphism and homology of HPV16 tumor-associated antigen (E6) gene in cervical cancer in Guangdong Province. Methods HPV163 genotypes were determined by universal primer PCR and direct sequencing. HPV16E6 was amplified by nested PCR from multiple HPV16-type specimens. The genetic variation was detected by DNA sequencing. Then analyze its homology. Results The detection rate of HPV-DNA in 50 cases of cervical cancer was 78%, including 18 cases of HPV16 and HPV18 mixed infection and 15 cases of HPV16 infection alone. HPV16E625 cases were amplified in 34 cases of HPV16 type specimens. Among them, 178 nucleotides showed a large variation with a mutation rate of 72%. The corresponding amino acids changed from aspartic acid to glutamic acid. Conclusion The region of base substitution of HPV16E6 DNA sequence is mainly located at amino terminal 94 ~ 241, and the carboxyl end is relatively conservative with no mutation. The hot spot mutation of HPV16E6 in cervical cancer in Guangdong is Nt178.