机构地区:[1]广州市番禺区中心医院消化中心,广东广州511400
出 处:《消化肿瘤杂志(电子版)》2024年第1期58-66,共9页Journal of Digestive Oncology(Electronic Version)
基 金:广州市科技计划项目(202102080538)。
摘 要:目的检测组蛋白伴侣抗沉默功能蛋白1B(anti-silencing functional protein 1B,ASF1B)在胃癌组织中的表达情况及其与胃癌患者预后之间的关系,探讨ASF1B对人胃癌细胞株AGS增殖、转移能力的影响以及可能的分子机制,分析ASF1B与幽门螺杆菌感染之间的关系。方法分析癌症基因组图谱数据库中泛瘤种的肿瘤组织和癌旁正常组织中ASF1B的表达情况,并通过实时荧光定量聚合酶链反应(real time fluorescent quantitative polymerase chain reaction,RT-qPCR)检测ASF1B在所收集的59例配对的胃癌组织及癌旁正常组织中的mRNA表达水平。分析Kaplan-Meier plotter数据库中胃癌患者ASF1B的表达水平与其预后之间的关系,后通过免疫组织化学检测胃癌组织芯片中ASF1B的表达水平并分析其与胃癌患者临床预后及临床病理参数之间的关系。通过小干扰RNA(small interfering RNA,siRNA)技术敲低胃癌细胞系AGS中的ASF1B表达,分组为siNC组(阴性对照转染细胞)、siASF1B#1组、siASF1B#2组及siASF1B#3组,后采用CCK-8、平板单克隆实验、Transwell小室迁移实验检测各组细胞的增殖、转移能力。分析基因表达综合数据库的数据集GSE27411中幽门螺杆菌阳性和阴性的胃癌组织中ASF1B的表达水平,并通过RT-qPCR及蛋白质印迹法检测ASF1B在幽门螺杆菌感染前后的胃癌细胞系AGS中的表达水平。结果相关数据库及我们的胃癌队列中的胃癌组织ASF1B的mRNA和蛋白表达水平均高于癌旁正常组织,且胃癌组织中高表达的ASF1B与胃癌患者不良预后相关。此外,与siNC组相比,敲低ASF1B组(siASF1B#1组、siASF1B#2组及siASF1B#3组)的细胞增殖和转移能力均减弱。最后,幽门螺杆菌阳性的胃癌组织及幽门螺杆菌感染后的人胃癌细胞AGS中的ASF1B表达水平均上调。结论ASF1B在幽门螺杆菌感染诱发的胃癌发生发展过程中可能起到重要的促进作用,且高表达的ASF1B与胃癌患者更早发生转移及较差�ObjectiveTo detect the expression level of histone chaperone anti-silencing functional protein 1B(ASF1B)in gastric cancer(GC)tissues and its relationship with the prognosis of GC patients.To investigate the effects of ASF1B on the proliferation and metastasis ability of human GC cell line AGS and the possible molecular mechanism.To analyze the relationship between Helicobacter pylori and the expression of ASF1B.Method The expression level of ASF1B in various of tumor tissues and adjacent normal tissues in the cancer genome atlas database was analyzed,and then the mRNA expression level of ASF1B in 59 pairs of GC tissues and paired adjacent normal tissues of our cohort were measured via real time fluorescent quantitative polymerase chain reaction(RT-qPCR).The relationship between the expression levels of ASF1B and the prognosis of GC patients in Kaplan-Meier plotter database was analyzed.Besides,the expression levels of ASF1B in GC tissue microarray were detected by immunohistochemistry,and then the relationship between ASF1B and clinical prognosis and clinicopathological parameters of GC patients was analyzed.Small interfering RNA(siRNA)technology was used to knockdown the expression of ASF1B in GC cell line AGS,and AGS cells were divided into siNC group(negative control transfected cells),siASF1B#1 group,siASF1B#2 group and siASF1B#3 group.Then the proliferation and metastasis ability of different groups were detected by CCK-8,plate monoclonal assay and Transwell cell migration assay.The expression levels of ASF1B in GC tissues with positive and negative Helicobacter pylori in gene expression omnibus database(GSE27411)were analyzed,and then the expression levels of ASF1B in GC cell line AGS before and after Helicobacter pylori infection were detected by RT-qPCR and western blot.Result The mRNA and protein expression levels of ASF1B in GC tissues of the database and our cohort were higher than those in adjacent normal tissues.Moreover,higher expression of ASF1B in GC tissues was correlated with the poorer prognosi
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