宫颈癌中HPV16病毒E6-295T/G和E7-647A/G多态性位点的致病特征及对免疫分子影响的机制研究

基于在新疆维吾尔族和汉族宫颈癌组织中HPV16病毒新发现的E6基因295T/G和E7基因647A/G多态性位点。本研究利用PCR结合基因测序方法分析宫颈鳞癌、腺癌及腺鳞癌组织中这些多态性位点的分布和致癌特征,并利用免疫组织化学实验检测宫颈组织中多态性位点对免疫分子表达的影响。利用基因转染实验将不同多态性位点克隆到真核表达载体，分析这些多态性位点对免疫相关分子的影响。利用裸鼠异种移植瘤方法，建立不同多态性位点的BALB/C裸鼠模型，观察成瘤体积和成瘤重量变化情况，检测天然免疫相关分子mRNA和蛋白质表达水平。分析多态性位点通过U-ISGF3复合物基因和“SUV39H1介导的甲基化沉默的免疫效应基因”途径对免疫分子表达影响的分子机制。阐明HPV16病毒E6-295T/G和E7-647A/G多态性位点对天然免疫分子表达的影响及分子机制,对于确定HPV16病毒风险、预后和防治宫颈癌有重要的意义。

It is based on the 295T/G polymorphism of E6 gene and 647A/G polymorphism of E7 gene in cervical cancer tissues of Uygur and Han nationalities in Xinjiang. In this study, PCR combined with gene sequencing was used to analyze the distribution and carcinogenic characteristics of these polymorphic sites in cervical squamous cell carcinoma, adenocarcinoma and adenosquamous carcinoma, and the effect of polymorphic sites on the expression of immune molecules in cervical tissue was detected by immunohistochemistry. Different polymorphic sites were cloned into eukaryotic expression vector by gene transfection experiment, and the effect of these polymorphic sites on molecules related to immunity was analyzed. BALB/C nude mice models with different polymorphic sites were established by xenotransplantation in nude mice. The volume and weight of tumor were observed, and the mRNA and protein expression levels of innate immune related molecules were detected. To analyze the molecular mechanism of the influence of polymorphic loci on the expression of immune molecules through the U-ISGF3 complex gene and "SUV39H1 mediated methylation silencing immune response gene" pathway. Elucidating the effect and molecular mechanism of HPV16 virus E6-295T/G and E7-647A/G polymorphisms on the expression of innate immune molecules is of great significance for determining HPV16 virus risk, prognosis, and preventing cervical cancer.