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微小 RNA-200b-3p 通过 C-X-C 基序趋化因子配体 12/CXC 趋化因子受体 7 轴抑制胃癌细胞增殖、迁移和侵袭。

MicroRNA-200b-3p restrains gastric cancer cell proliferation, migration, and invasion via C-X-C motif chemokine ligand 12/CXC chemokine receptor 7 axis.

机构信息

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.

出版信息

Bioengineered. 2022 Mar;13(3):6509-6520. doi: 10.1080/21655979.2022.2034585.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d79/8974025/cb73150edc22/KBIE_A_2034585_UF0001_OC.jpg

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