Objective To investigate the relationship between the Fas-associated factor 1 (FAF1)expression with Helicobacter pylori (Hp) infection and carcinogenesis of gastric cancer.Methods 8 × 105 cells of human gastric carcinoma cells (HGC-27) and human gastric epithelial cells (GES-1) were co-cultured with Hp standard strain NCTC11637 for 24 h,and the cells were divided by cell/bacterial ratio into three groups:control group,1∶100 and 1∶200 co-culture groups.Each experiment was conducted three times and each sample was assayed in triplicate.Thiazolyl blue (MTT) assay was used to determine the proliferation in 490 nm,flow cytometry to determine the apoptosis of 1 × 104 cells per group,and real-time fluorescent quantitative reverse transcriptase polymerase chain reaction (FQ-PCR) to detect the relative expression of FAF1 mRNA.The sequence length was 164 bp.Results After co-culture with Hp,proliferation of cells was decreased and apoptosis was increased.As compare with respective control group,the relative expression of FAF1 mRNA in 1 ∶ 100 co-culture group and 1 ∶ 200 co-culture group in HGC-27 cells was significantly decreased (0.67 ± 0.20 vs.0.99 ± 0.27,F =11.51,P ＜ 0.05 ; 0.44 ± 0.30 vs.0.99 ±0.27,F =11.5 1,P ＜ 0.01).The relative expression of FAF1 mRNA in 1∶100 co-culture group in GES-1 cells was significantly increased (1.61 ± 0.77 vs.0.99 ± 0.41,F =14.45,P ＜ 0.05),and that in 1∶200 co-culture group in GES-1 cells was significantly decreased (0.36 ±0.22 vs.0.99 ±0.41,F =14.45,P ＜0.05).As compared with control group,1∶100 and 1∶200 co-culture groups of GES-1 cells,the expression levels of FAF1 mRNA in each corresponding group of HGC-27 cells were significantly decreased (0.33±0.16vs.0.99±0.41,t=4.74,P＜0.01; 0.17±0.12 vs.1.08±±0.53,t=5.28,P＜0.01;0.39±0.24 vs.0.98 ±0.23,t=5.67,P＜0.01).Conclusion Hp infection can down-regulate the expression of FAF1 gene,leading to gastric cell proliferation and apoptosis imbalance,then inducing the carcinogenesis of gastric cancer.%目的 探讨Fas相关因子l(FAF1)基因表达与幽门螺杆菌感染(Hp)及胃癌发生之间的关系.方法 人胃癌细胞株HGC-27、人胃黏膜上皮细胞GES-1 8×105个与Hp标准菌株NCTC11637共培养24h,按细胞/细菌比分为3组:空白对照组、1∶100和1∶200共培养组,每组实验设3个复孔,采用噻唑蓝(MTT)比色法490 nm波长处测量吸光度值计算细胞存活率,流式细胞仪检测l×104个细胞凋亡率,荧光实时定量聚合酶链反应(FQ-PCR)检测细胞FAFl mRNA相对表达量,扩增片段长度为164 bp.结果 HGC-27和GES-1细胞与Hp共培养后,细胞增殖均减少,凋亡率增加.与各自空白对照组比较,FAFl mRNA在HGC-27细胞1∶100和1∶200共培养组表达量明显降低(0.67 ±0.20比0.99 ±0.27,F=11.51,P＜0.05;0.44 ±0.30比0.99±0.27,F=11.51,P＜0.01);FAFl mRNA在GES-1细胞1∶100共培养组表达量明显升高(1.61 ±0.77比0.99 ±0.41,F=14.45,P＜0.05),1∶200共培养组表达量明显降低(0.36 ±0.22比0.99±0.41,F=14.45,P＜0.05).与GES-1细胞空白对照组、1∶100和1∶200共培养组比较,FAF1 mRNA在HGC-27相应各组表达量均明显降低(0.33 ±0.16比0.99 ±0.41,t =4.74,P＜0.01;0.17±0.12比1.08 ±0.53,t=5.28,P＜0.01;0.39 ±0.24比0.98±0.23,t=5.67,P＜0.01).结论 Hp感染下调FAF1基因表达,导致细胞增殖和凋亡失衡,诱发胃癌的发生.
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