Department of General Gastropathy, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
Department of Gastrosurgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
Bioengineered. 2022 Mar;13(3):6509-6520. doi: 10.1080/21655979.2022.2034585.
This study was conducted to investigate the impact of microRNA (miR)-200b-3p on viability, migration, and invasion of gastric cancer (GC) cells and its mechanism. Quantitative real-time PCR (qRT-PCR) was conducted to measure miR-200b-3p expression in GC tissues and cells; besides, the relationship between miR-200b-3p expression and overall survival time (OS) was analyzed with OncomiR database; cell counting kit-8 (CCK-8), colony formation assay, flow cytometry, scratch healing assay, and Transwell assay were performed to detect the proliferation, cell cycle progression, migration, and invasion of GC cells; a lung metastasis model in nude mice was used to examine the effect of miR-200b-3p on the metastasis of GC cells ; the interplay between miR-200b-3p and C-X-C motif chemokine ligand 12 (CXCL12) mRNA 3' UTR was predicted by bioinformatics and verified with a dual-luciferase reporter gene assay; besides, the expression of CXCL12 and CXC chemokine receptor 7 (CXCR7) was probed by Western blot. It was found that miR-200b-3p expression was down-regulated in GC tissues, which was remarkably associated with the lymph node metastasis and decrease of differentiation of GC; transfection with miR-200b-3p mimics restrained the growth, migration, and invasion of GC cells , induced cell cycle arrest, and inhibited CXCL12 and CXCR7 expression levels; transfection of miR-200b-3p inhibitors worked oppositely and promoted lung metastasis . CXCL12 was confirmed as the downstream target of miR-200b-3p and was negatively modulated by miR-200b-3p. In conclusion, miR-200b-3p inhibited GC progression via regulating CXCL12/CXCR7 axis.
本研究旨在探讨 microRNA (miR)-200b-3p 对胃癌 (GC) 细胞活力、迁移和侵袭的影响及其机制。采用实时定量聚合酶链反应 (qRT-PCR) 检测 GC 组织和细胞中 miR-200b-3p 的表达;此外,利用 OncomiR 数据库分析 miR-200b-3p 表达与总生存时间 (OS) 的关系;采用细胞计数试剂盒-8 (CCK-8)、集落形成实验、流式细胞术、划痕愈合实验和 Transwell 实验检测 GC 细胞的增殖、细胞周期进程、迁移和侵袭;建立裸鼠肺转移模型检测 miR-200b-3p 对 GC 细胞转移的影响;通过生物信息学预测 miR-200b-3p 与 C-X-C 基序趋化因子配体 12 (CXCL12) mRNA 3'UTR 的相互作用,并通过双荧光素酶报告基因实验进行验证;此外,采用 Western blot 检测 CXCL12 和 CXC 趋化因子受体 7 (CXCR7) 的表达。结果发现,miR-200b-3p 在 GC 组织中表达下调,与 GC 的淋巴结转移和分化程度降低显著相关;转染 miR-200b-3p 模拟物抑制 GC 细胞的生长、迁移和侵袭,诱导细胞周期停滞,并抑制 CXCL12 和 CXCR7 表达水平;转染 miR-200b-3p 抑制剂则作用相反,促进肺转移。CXCL12 被证实是 miR-200b-3p 的下游靶基因,并受 miR-200b-3p 负调控。综上所述,miR-200b-3p 通过调节 CXCL12/CXCR7 轴抑制 GC 进展。