局部细胞中HPVl6基因表达量及突变与其宫颈持续感染的关系

目的探讨宫颈脱落细胞中人乳头瘤病毒（HPV）16基因的表达量及突变与其宫颈持续感染的关系。方法选取96例HPVl6感染患者，其中33例宫颈脱落细胞HPVl6持续阳性（持续感染组），63例6个月后复查转为阴性（非持续感染组）。采用半定量PCR技术检测两组（宫颈脱落细胞中）HPVl6E5、E6、E7及长控制区（LCR）基因表达量，基因测序法检测HPVl6E5、E6、E7及LCR基因突变。结果持续感染组HP~16E5、E6、E7、LCR基因表达量分别为0．20±0．02、0．30±0．02、0．19±0．01、0．53±0．02，非持续感染组分别为0．17±0．02、0．27±0．03、0．16±O．06、0．504-0．04，两组比较P均〉0．05。持续感染组与非持续感染组3978A→c（144L）位点突变例数分别为33、63例，4041A→G（165V）位点突变例数分别为33、63例，4076A—T（同义突变）位点突变例数分别为28、31例；两组4076A→T突变例数比较，P〈0．01。持续感染组与非持续感染组E6基因178T→G（D25E）位点突变例数分别为10、4例，两组比较，P〈0．05；持续感染组与非持续感染组E7基因647A→G（N29S）与846T→c（同义突变）联合突变例数分别为9、8例，两组比较，P〉0．05；持续感染组与非持续感染组7761c→T位点突变例数分别为33、63例，P〉0．05；7727A→c位点突变例数分别为14、4例，两组比较，P〈0．01。结论宫颈脱落细胞中HPVl6基因表达量与持续性感染无密切关系，HPVl6E5、E6、LCR基因突变与宫颈持续性感染有关，其中HPVl6E54076A→T（同义突变）、E6178T→G（D25E）与LCR7727A→c突变点可能是导致HPVl6持续感染的关键突变位点。

Correlations of HPV16 gene expression and mutation in local cells with cervical virus persistent infection

Objective To explore the correlation between the HPV16 gene in cervical exfoliated cells and cervical HPV16 persistent infection. Methods A total of 96 patients with cervical exfoliated cells were collected. After 6 months observation, among them, 33 cases were detected HPV16 persistent positive （persistent infection group）, and the other 63 cases turned to be negative （non-persistent infection group）. The expression and mutation of DNA in HPV16 ES, E6, E7 and long centralization control region （LCR） were detected by semi-quantitative PCR technology and gene sequencing re- spectively. Results DNA expression of HPV16 ES, E6, E7, LCR were 0.20 ± 0.02, 0.30 ± 0.02, 0.19 ± 0.01, and O. 53 ±： O. 02 in the persistent infection group ; and 0.17 ± 0.02, 0.27 ± O. 03, 0.16 ±0.06, 0.50 ± 0.04 in the non-per- sistent infection group （ all P 〉 0.05 ）. For E5 gene, in the two groups, 33 and 63 were detected with mutation of 3978A→ C（I44L） ; 33 and 63 with mutation of 4041A→G （I65V） ; 28 and 31 with mutation of 4076A→T （ P 〈 0. O1 ）. For E6 gene, 10 and 4 were detected with mutation of 178T→G （D25E） （P 〈0.05）. For E7 gene, 9 and 8 were detected with mutation of 647A→G（N29S） and 846T→C （P 〉 0.05）; 33 and 63 were detected with mutation of 7761C→T （P 〉 0.05 ） ; 14 and 4 were detected with mutation of 7727A→C （ P 〈 0.01 ）. Conclusions The expression of HPV16 DNA in cervical exfoliated cells may not be relevant to HPV16 persistent infection, but the internal genetic mutations of HPV16 ES, E6 and LCR may contributed to it. The following mutations, HPV16 E5 4076 A→T （synonymous mutations）, E6 178 T→ G （D25E） and LCR 7727A→C may be the key factors that lead to HPV16 persistent infection.