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目的探讨丙型肝炎病毒(hepatitis C virus,HCV)F蛋白与多发性腺瘤样息肉病(adenomatous polyposis coli,APC)基因甲基化在慢性HCV感染中的关系。方法收集慢性HCV患者(chronic hepatitis patient,CHP)的血样,检测F抗体(F antibody,F-Ab)的阳性率并且分成两组(F-Ab(+)CHP组、F-Ab(-)CHP组),收集健康者的血样作为对照组;分离外周血单核细胞(peripheral blood mononuclear cells,PBMCs),分别用HCV Core/F蛋白刺激,72 h后,分别提取DNA,用二代测序检测APC基因的甲基化水平,分析各组APC基因甲基化程度的差异。结果APC基因甲基化程度在F-Ab(+)CHP组、F-Ab(-)CHP组及健康对照组三组间差异有统计学意义(F=185.185,P<0.001),F-Ab(+)CHP组APC基因甲基化程度高于F-Ab(-)CHP组,健康对照组最低(均有P<0.05);三组样本PBMC分别经Core/F蛋白体外刺激后,APC基因甲基化程度各组总体比较差异均有统计学意义(均有P<0.05),Core蛋白与F蛋白共刺激组细胞APC基因甲基化程度高于F蛋白刺激组,Core蛋白刺激组为最低。结论在慢性HCV感染患者中,HCV F蛋白的产生能够影响APC基因甲基化程度。
Objective To investigate the relationship between hepatitis C virus (HCV) F protein and methylation of multiple adenomatous polyposis coli (APC) gene in chronic HCV infection. Methods The blood samples of chronic hepatitis patients (CHP) were collected and the positive rate of F antibody (F-Ab) was detected and divided into two groups (F-Ab (+) CHP group, F-Ab The peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with HCV Core / F protein respectively. After 72 hours, the DNA was extracted separately and the second generation sequencing was used to detect the APC The methylation level of APC gene was analyzed. The difference of methylation level of APC gene in each group was analyzed. Results The methylation level of APC gene was significantly different among F-Ab (+) CHP group, F-Ab (-) CHP group and healthy control group (F = 185.185, P <0.001) The level of APC gene methylation in CHP group was higher than that in F-Ab CHP group (all P <0.05). The PBMCs of three groups were stimulated by Core / F protein in vitro and APC gene The degree of methylation was significantly higher in all groups (P <0.05). The methylation level of APC gene in cells co-stimulated with Core protein and F protein was higher than that in F protein stimulation group, and Core protein stimulation group was the lowest . Conclusion HCV F protein production can affect the methylation level of APC gene in patients with chronic HCV infection.