ARMC8在胃癌细胞恶性进展中的机制研究

Mechanistic study of ARMC8 in malignant progression of gastric cancer cells

目的 观察胃癌(gastric cancer,GC)组织、血浆和细胞系中ARMC8(armadillo repeat containing protein 8)的表达水平,探究ARMC8对GC细胞侵袭、迁移及上皮间充质转化(epithelial-mesenchymal transition,EMT)的分子机制。方法 通过GEPIA数据库对408例GC组织和211例正常胃组织中的ARMC8的表达进行差异分析;通过实时荧光定量聚合酶链反应(quantitative real-time PCR,qRT-PCR)检测ARMC8在GC患者血浆及正常人血清的相对表达量;在2种GC细胞系(HGC-27、AGS)和1种正常胃黏膜上皮细胞(GES-1)中进一步验证ARMC8的表达情况。通过转染siARMC8及阴性对照实现下调GC细胞中ARMC8的表达水平。随后,采用Transwell细胞侵袭实验、伤口愈合实验检测GC细胞的侵袭和迁移能力;用WB检测下调ARMC8的表达对EMT相关蛋白及Wnt/β-catenin信号通路的影响。结果 ARMC8在GC组织、血浆及GC细胞系中均为高表达。Transwell侵袭实验和伤口愈合实验显示,下调ARMC8抑制了GC细胞的侵袭和迁移能力。此外,下调ARMC8抑制了GC细胞EMT过程及Wnt/β-catenin信号通路的激活。结论 ARMC8在GC组织、血浆及细胞系中均呈高表达,下调ARMC8可能通过抑制Wnt/β-catenin信号通路的激活来抑制GC的侵袭、迁移和EMT过程。

Objective In order to observe the expression level of armadillo repeat containing protein 8(ARMC8)in gastric cancer(GC)tissues,plasma and cell lines,and to explore the molecular mechanism of ARMC8 on invasion,migration and epithelialmesenchymal transition(EMT)of GC cells.Methods The differential expressions of ARMC8 in 408 GC tissues and 211 normal tissues were analyzed from GEPIA database.The expression of ARMC8 in the plasma of GC patients and healthy people was analyzed by quantitative realtime PCR(qRT-PCR).The expression of ARMC8 was further validated in two GC cell lines(HGC-27,AGS)and one normal gastric mucosal epithelial cell(GES-1).The expression level of ARMC8 was downregulated by transfection with si-ARMC8 and negative control(NC).The abilities of migration and invasion of GC cells were determined by wound healing assay and Transwell assay after transfection.The effect of downregulation of ARMC8 expression on EMT-associated proteins and the activation of Wnt/β-catenin signaling was measured by WB.Results The expression of ARMC8 was significantly elevated in GC tissues plasma and GC cell lines;Transwell invasion assay and wound healing results showed that downregulation of ARMC8 inhibited GC cell invasion and migration ability.Downregulation of ARMC8 inhibited the EMT process and the activation of Wnt/β-catenin signaling pathway in GC cells.Conclusion ARMC8 is highly expressed in GC tissues,the plasma of GC patients and GC cell lines,and which may play a key role in migration invasion and EMT of GC via Wnt/β-catenin signaling.