宫颈癌中HPV感染分型与EGFR靶向治疗药物敏感突变、EGFR启动子甲基化的研究

Relationship among HPV Genotypes, EGFR Promoter Methylation and EGFR Mutations in Cervical Cancer

目的检测并分析宫颈癌组织标本中感染HPV病毒型别与EGFR基因启动子甲基化状态、EGFR靶向治疗药物敏感相关基因突变状态的分布与关联性，为指导宫颈癌的个体化治疗提供新思路和分子诊断技术方法。方法分别提取180例宫颈癌组织DNA，GP5＋／GP6＋非对称扩增荧光偏振检测方法检测每例宫颈癌标本组织DNA的HPV16、18、58、52型。非对称扩增荧光偏振检测方法检测每例宫颈癌标本组织的亚硫酸氢盐转变DNA的EGFR启动子甲基化状态：测序法检测每例宫颈癌标本组织DNA的EGFR基因18、19、21外显子突变状态；统计分析检On,0结果。结果本研究成功检测了宫颈癌标本组织DNA中HPV16、18、58、52型和EGFR基因18、19、21外显子突变状态，成功检测了宫颈癌标本组织亚硫酸氢盐转变DNA的EGFR启动子甲基化状态。EGFR启动子甲基化阳性65例，阳性率为36．1％。单纯HPV16阳性54例，其中EGFR启动子甲基化34例，占63．0％。HPV16阳性样本中EGFR启动子甲基化阳性比率最高。结论本研究中，宫颈癌标本中HPV感染率为100％，宫颈癌标本中未发现EGFR药物敏感相关基因EGFR18、19、21外显子突变，宫颈癌标本中EGFR启动子甲基化阳性率较高，其中，HPV16感染的宫颈癌标本中EGFR启动子甲基化阳性率较其他病毒型显著增高，提示HPV16感染与EGFR启动子甲基化有一定关联．

Objective To examine the distribution and relationship among HPV genotypes, EGFR promoter methylation and EGFR mutations in cervical cancer in order to provide a novel un-derstanding to this disease and an applicable pharmacogenomic tool for individualized management of cervical cancer patients. Methods DNA was extracted from 180 tissue samples of cervical cancer. HPV genotypes （HPV16, 18, 58, 52） were detected in these tissue samples by asymmetric GP5 ＋/6 ＋ PCR and hybridization with fluorescence polarization assay. EGFR promoter methylation was detected by asymmetric PCR and hybridization with fluorescence polarization assay. The mutations of EGFR exons 18, 19, 21 were detected by sequencing. Results Sixty-five of 180 cervical cancer samples were methylated and the methylation rate was 36. 1% , and all the 180 cervical cancer sam-ples were HPV positive, among which 54 were HPV16 positive. Thirty-four of the 54 HPV16-posi-rive samples had the methylation of EGFR promoter. No EGFR exons 18, 19, 21 mutation was de-tected. Conclusion HPV16-positive samples of cervical cancer had higher methylation of EGFR promoter than samples positive for other HPV types, suggesting close association of HPV infection with EGFR promoter mehtylation.