目的 探讨宫颈鳞状细胞癌HPV16 E6、E7基因突变的位点及发生频率.方法 前瞻性研究宫颈鳞状细胞癌标本108例,癌临床分期I期44例,Ⅱ-Ⅲ期39例,Ⅳ期25例;肿瘤分化为中高分化的87例,低分化21例;有淋巴结转移的19例,无淋巴结转移的89例.均确认为HPV16感染阳性标本,针对HPV16 E6、E7基因设计合成引物、PCR扩增、产物DNA纯化、测序、基因突变检测.结果 HPV16 E6 T178A/G和HPV16 E7A647G突变是HPV16 E6、E7基因的常见突变位点,突变例数分别为46例和63例,突变率为42.6％和63.9％,这两个突变位点的突变率分别与是否有淋巴结转移和肿瘤分化程度相关(P＜0.05),与肿瘤病理分期不相关(P＞0.05).结论 HPV16 E6 T178A/G和HPV16 E7A647G突变与是否有淋巴结转移和肿瘤分化程度有关,其相关检测可作为提示及评估宫颈癌严重程度的指标.%Objective To explore the mutation site and frequency of HPV16 E6 and E7 in cervical carcinoma. Methods A total of 108 specimens of cervical squamous cell carcinoma were included in this prospective study, among them 44 specimens were at clinical stage 1,39 cases were at stage Ⅱ- Ⅲ and 25 cases were at stage Ⅳ. The medium and high differentiation had been seen in 87 cases and 21 cases were poorly differentiated. There were 19 cases with metastasis in lymph nodes, and 89 cases without metastasis in lymph nodis. All of them were detected as HPV16 positive samples. Design and synthetic primers of HPV16 E6 and E7 genes had been established. After PCR amplification,product purification,sequencing and gene mutation detection was executed. Results Common mutations in HPV16 E6 and E7 genes were E6 T178A / G and E7 A647G. The number of mutations was 46 and 63 respectively. Their mutation rates were 42.6％ and 63.9％ respectively. They were associated with metastasis in lymph nodes and tumor differentiation (P ＜0.05), but they did not correlate with tumor staging. Conclusion HPV16 E6 T178A/G and HPV16 E7 A647G mutations are associated with metastasis in lymph nodes and tumor differentiation. It can be used as an indicator for the severity of cervical cancer.
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