宫颈病变中HPV16感染状况、微卫星不稳定性及错配修复基因表达的研究

目的通过检测HPV16、微卫星不稳定性（microsatellitein stability,MSI）和错配修复基因（hMLH1、hMSH2）的表达,探讨三者在宫颈病变进展中的作用及意义。方法将宫颈病变组织分为宫颈炎、宫颈上皮内瘤变（cervicalintraepithelialneoplasia,CIN）CIN1、CIN2～CIN3、宫颈癌4组。用PCR方法检测HPV16感染状况,用聚合酶链-单链构象多态性分析方法检测宫颈组织中MSI的表达,采用免疫组织化学SP法来检测宫颈组织中的hMLH1和hMSH2的表达,并分析三者的相关性。结果CIN120例中25.0%（5/20）检测到HPV16,CIN2～CIN331中54.8%（17/31）检测到HPV16,二者比较χ2=4.413,P=0.034;宫颈癌中各期HPV16感染比较,差异均无统计学意义。选取的三个微卫星位点D3S2832E、RH91127和SHGC-56838均在宫颈炎中未检测到MSI,在CIN和宫颈癌中均检测到MSI的存在。但总体的表达率较低,最高只有46.7%。错配修复基因的低表达检测结果和MSI的检测结果一致,唯一不同的是前者的总体表达率较高,最高的低表达率为80%;宫颈病变中MSI与错配修复基因（本文即hMLH1、hMSH2）蛋白低表达及HPV16为正相关。结论HPV16的感染导致错配修复基因的低表达,进一步引起微卫星不稳定性,可能是宫颈癌发生、发展的重要机制,HPV16、微卫星不稳定性和错配修复基因有望作为宫颈癌高危人群的检测指标。

The study of HPV16 infection,microsatellite instability and mismatch repair gene expression in cervical lesion pathological changes

Objective Our objective was to explore the importance of human papilloma viru 16(HPV16) in the etiology of cervical intraepithelial neoplasia(CIN) by analyzing the expression of human mut-l homologuel,human mut-s homologue2 and microsatellite instability (MI)in patients suffering from cervical lesion.Methods Patients were divided into 4 groups according to the pathologic diagnosis:cervicitis,CIN1,CIN2～CIN3 and cervical cancer groups.MI was determined by polymerase chain reaction single strand conformation polymorphismanalysis (PCR-SSCP).Immunohistochemical staining with SP were performed to analyse the gene expression of human mut-l homologuel and human mut-s homologue2(hMLH1 and hMSH2),also PCR was performed to detect human papilloma viru 16.Results 5 pantiens were HPV16 positive in CIN1 group (5/20,25.0%,) which was significant lower than that in CIN1 group(17/31,54.8%;χ2=4.413,P=0.034).The selected three MSI loci D3S2832E,RH91127 and SHGC-56838 were not detected in cervicitis group while detected in both CIN and cervical cancer group,but the general expression rate was low (46.7%).And the expression of hMLH1and hMSH2 had the same trend with MSI and were positive correlated with HPV16 infection.Conclusions HPV16 infection can decrease the expression of hMLH1 and hMSH2 and thus cause MSI which may be an important mechanism of the development of cervical cancer.HPV16 infection,hMLH1 and hMSH2 MSI is expected to be makers of high-risk of cervical carcinoma.