摘要
目的分析敲低SH2结构域蛋白2A(SH2D2A)基因对胃癌细胞生物学行为的影响。方法从癌症基因组图谱数据库获取胃癌样本RNAseq数据,分析SH2D2A基因在胃癌组织及癌旁正常组织中的表达。通过慢病毒转染技术构建SH2D2A基因沉默表达的慢病毒载体,将人胃癌细胞AGS分为shCtrl组(转染慢病毒空载体)和shSH2D2A组(转染SH2D2A基因沉默表达的慢病毒载体),采用实时荧光定量PCR和Western blot检测SH2D2A基因敲低效果。采用Celigo细胞计数法、AnnexinⅤ-APC单染法流式细胞仪、Transwell实验分别检测两组细胞的增殖、凋亡、迁移及侵袭能力。采用Western blot检测敲低SH2D2A基因对丝氨酸/苏氨酸蛋白激酶(AKT)、磷酸化AKT(p-AKT)、v-Myc骨髓细胞瘤病毒癌基因同源物(c-MYC)蛋白表达的影响。结果SH2D2A基因在胃癌组织中的表达水平高于癌旁正常组织(P<0.05)。与shCtrl组相比,shSH2D2A组的SH2D2A mRNA及蛋白表达水平降低,读板第2至第5天时胃癌细胞数量减少,胃癌细胞凋亡率升高,迁移细胞数量及侵袭细胞数量减少,AKT、p-AKT、c-MYC蛋白表达水平降低(P<0.05)。结论SH2D2A基因在胃癌组织中高表达,沉默SH2D2A基因可抑制胃癌细胞增殖、迁移及侵袭能力,并促进其凋亡,该作用机制可能与其调控AKT信号通路及c-MYC信号通路共同发挥对胃癌的抑制效应有关。
Objective To analyze the effect of knocking down SH2 domain containing 2A(SH2D2A)gene on biological behavior of gastric cancer cells.Methods RNAseq data of gastric cancer samples were obtained from The Cancer Genome Atlas database to analyze the expression of SH2D2A gene in gastric cancer tissues and para-carcinoma normal tissues.A lentiviral vector for silencing SH2D2A gene expression was constructed using lentiviral transduction technology.Human gastric cancer AGS cells were divided into shCtrl group(transfected with an empty lentiviral vector)or shSH2D2A group(transfected with the SH2D2A-silencing lentiviral vector).The SH2D2A gene knockdown effect was verified by real-time fluorescent quantitative PCR and Western blot.Celigo cell counting assay,AnnexinⅤ-APC single staining flow cytometry,and Transwell assays were used to detect cell proliferation,apoptosis,migration,and invasion capabilities,respectively.Western blot was employed to detect the effects of SH2D2A gene knockdown on the protein expressions of serine/threonine protein kinase(AKT),phosphorylated AKT(p-AKT),and v-Myc myelocytomatosis viral oncogene homolog(c-MYC).Results The expression of SH2D2A gene was higher in gastric cancer tissues than in para-carcinoma normal tissues(P<0.05).Compared with the shCtrl group,the shSH2D2A group exhibited decreased mRNA and protein expressions of SH2D2A,reduced gastric cancer cell counts from days 2 to 5 of plate reading,increased gastric cancer cell apoptosis rate,decreased numbers of migrated and invaded cells,and reduced protein expressions of AKT,p-AKT and c-MYC(P<0.05).Conclusion SH2D2A gene is highly expressed in gastric cancer tissues.Silencing SH2D2A gene can inhibit the proliferation,migration,and invasion capabilities of gastric cancer cells and promote their apoptosis.This mechanism may be related to its regulation of the AKT signaling pathway and the c-MYC signaling pathway,collectively exerting inhibitory effects on gastric cancer.
作者
黄彩家
陈智斌
黄名威
王婷安
韦尉元
HUANG Caijia;CHEN Zhibin;HUANG Mingwei;WANG Ting′an;WEI Weiyuan(Department of Gastrointestinal Surgery,Guangxi Medical University Cancer Hospital,Nanning 530021,Guangxi,China)
出处
《广西医学》
2025年第12期1789-1795,共7页
Guangxi Medical Journal
基金
广西自然科学基金(2021GXNSFAA220036、2024GXNSFAA010407、2023GXNSFAA026133)。
关键词
胃癌
SH2结构域蛋白2A
细胞凋亡
细胞增殖
细胞迁移
细胞侵袭
Gastric cancer
SH2 domain containing 2A
Cell apoptosis
Cell proliferation
Cell migration
Cell invasion
