HPV16 E6-295T/G、E7-647A/G多态性位点在子宫颈鳞状细胞癌中的分布及对IRF9的影响

Distribution of HPV16 E6-295T/G and E7-647A/G polymorphisms in cervical squamous cancer and their effects on IRF9

目的分析HPV16 E6-295T/G、E7-647A/G多态性位点在子宫颈鳞状细胞癌、CIN1中的分布及其在维吾尔族与汉族之间分布的特点,探讨其对IRF9表达的影响。方法运用TCGA数据库及GEPIA数据库分析子宫颈癌组织中IRF9的表达及其与子宫颈癌肿瘤分级的关系;收集67例维吾尔族子宫颈鳞状细胞癌标本、21例维吾尔族CIN1标本,68例汉族子宫颈鳞状细胞癌标本和28例汉族CIN1标本,从收集的标本中提取DNA,PCR扩增E6和E7基因,PCR产物纯化后测序。将HPV16感染阳性标本进行免疫组化检测。结果维吾尔族子宫颈鳞状细胞癌标本中E6-295T/G、E7-647A/G变异分别为31例(46.27%)和7例(10.45%),CIN1标本中变异均为7例(33.33%)。汉族子宫颈鳞状细胞癌标本中E6-295T/G、E7-647A/G变异分别为12例(17.65%)和37例(54.41%),CIN1变异分别为8例(28.57%)和13例(46.43%)。维吾尔族子宫颈鳞状细胞癌中E6-295T/G的变异率高于汉族,汉族子宫颈鳞状细胞癌中E7-647A/G的变异率高于维吾尔族,差异有统计学意义(P<0.05)。CIN1标本中两民族间E6-295T/G和E7-647A/G的变异,差异无显著性(P>0.05)。免疫组化结果显示,IRF9在子宫颈鳞状细胞癌中的表达高于CIN1(P<0.05),子宫颈鳞状细胞癌标本中E6-295G/E7-647A变异组IRF9的表达高于E6-295T/E7-647G变异组和E6-295T/E7-647A未变异组(P<0.05)。E6-295T/E7-647G变异组与E6-295T/E7-647A未变异组的IRF9蛋白表达差异无显著性(P>0.05)。结论维吾尔族子宫颈鳞状细胞癌中HPV16 E6-295T/G变异高于汉族,汉族E7-647A/G变异高于维吾尔族。HPV16 E6-295T/G变异的子宫颈鳞状细胞癌组织中IRF9蛋白表达水平增高,可能在子宫颈癌的发生中起作用。

Purpose To study the distribution of polymorphic loci E6-295T/G and E7-647A/G of human papillomavirus type 16 in cervical squamous cell carcinoma and CIN1 in Uyghur and Han nationalities,respectively,and to explore the effects of their expression on natural immune molecules interferon regulatory factor 9(IRF9).Methods UALCAN database and GEPIA database were used to analyze the expression of IRF9 in cervical cancer and its relationship with tumor grade in cervical cancer patients.Then paraffin-embedded tissue samples of cervical cancer of Han and Uygur were collected,and DNA was extracted from 67 cervical squamous cell carcinoma of Uygur specimens,21 CIN1 of Uygur specimens,68 cervical squamous cell carcinoma of Han specimens,and 28 Han CIN1 specimens.E6 and E7 genes were amplified by PCR,and PCR products were purified and sequenced.The samples with positive HPV16 infection were selected and immunohistochemical experiments were performed to analyze the effects of polymorphic loci on natural immune molecules IRF9.Results In Uygur group,the variation of E6-295T/G and E7-647A/G were 31(46.27%)and 7(10.45%)in cervical squamous cell carcinomas,and the variation was 7(33.33%)and 7(33.33%)in CIN1,respectively.In the Han group,the variation of E6-295T/G and E7-647A/G were 12(17.65%)and 37(54.41%)in cervical squamous cell carcinomas,and the variation was 8(28.57%)and 13(46.43%)in CIN1,respectively.The variation rate of E6-295T/G in cervical squamous cell carcinoma of Uygur was higher than that in Han,and E7-647A/G in cervical squamous cell carcinoma of Han was higher than that in Uygur,the difference is statistically significant.However,there was no difference in the variation between the two ethnic groups in CIN1 by Chi-square test(P>0.05).In addition,immunohistochemical results showed that the expression level of IRF9 molecule in squamous cell carcinoma was higher than that in CIN1 group(P<0.05),and in all squamous cell carcinoma samples,the expression level of IRF9 molecule in E6-295G/E7-647A variant group was higher than that in E6-295T/E7-647G variant group and E6-295T/E7-647A group(P<0.05).There was no difference in IRF9 protein expression between E6-295T/E7-647G and E6-295T/E7-647A(P>0.05).Conclusion The variation of HPV16 virus E6-295T/G in Uyghur is higher than that in Han,and the variation of HPV16 virus E7-647A/G in Han is higher than that in Uygur.The increased expression of IRF9 protein in cervical squamous cell carcinoma with HPV16 virus E6-295T/G variation may play a role in the development of cervical cancer.