Biomed Pharmacother. 2017 Dec;96:782-791. doi: 10.1016/j.biopha.2017.08.144. Epub 2017 Nov 6.
This study intended to explore the molecular mechanism of RUNX3 in inhibiting the process of migration and proliferation of gastric cancer (GC) cells.
The overexpressed plasmids of RUNX3 and the interfering siRNA of RUNX3 were transfected into GC cells. Then, qRT-PCR and western blot were performed to identify the expression level of RUNX3 and miR-182 in tumors and adjacent tissues respectively. ChIP and luciferase assay were performed to detect the relationship between RUNX3 and miR-182 as well as miR-182 and HOXA9. Furthermore, EdU assay were used to investigate the proliferation of GC cells, Transwell assay and wound healing assay were utilized to assess cell metastasis. Xenograft mouse model was set to evaluate the proliferation in vivo.
The results of qRT-PCR and western blot indicated that RUNX3 could regulate the expression of miR-182. RUNX3 can be straightly interacted with the promoter region of miR-182 in accordance with the results of ChIP. Luciferase assay revealed that HOXA9 was the direct target gene of miR-182. In addition, EdU proliferation, wound healing assay and transwell assay showed that miR-182 mimics and HOXA9 siRNA could inhibit the ability of cells proliferation, migration and invasion. The findings of in vivo experiments strongly supported the view that miR-182/HOXA9 was involved in the process of RUNX3-mediated GC tumor growth.
RUNX3 could impede the ability of GC cells proliferation, migration and invasion by modulating miR-182/HOXA9 pathway.
本研究旨在探讨 RUNX3 在抑制胃癌（GC）细胞迁移和增殖过程中的分子机制。
将 RUNX3 的过表达质粒和 RUNX3 的干扰 siRNA 转染到 GC 细胞中。然后，通过 qRT-PCR 和 Western blot 分别鉴定肿瘤和相邻组织中 RUNX3 和 miR-182 的表达水平。通过 ChIP 和荧光素酶测定检测 RUNX3 和 miR-182 以及 miR-182 和 HOXA9 之间的关系。此外，通过 EdU 测定法研究 GC 细胞的增殖，通过 Transwell 测定法和划痕愈合测定法评估细胞转移。建立异种移植小鼠模型以评估体内增殖。
qRT-PCR 和 Western blot 的结果表明，RUNX3 可以调节 miR-182 的表达。根据 ChIP 的结果，RUNX3 可以与 miR-182 的启动子区域直接相互作用。荧光素酶测定显示，HOXA9 是 miR-182 的直接靶基因。此外，EdU 增殖、划痕愈合测定法和 Transwell 测定法表明，miR-182 模拟物和 HOXA9 siRNA 可以抑制细胞增殖、迁移和侵袭的能力。体内实验的结果有力地支持了 miR-182/HOXA9 参与 RUNX3 介导的 GC 肿瘤生长过程的观点。
RUNX3 通过调节 miR-182/HOXA9 通路可以抑制 GC 细胞的增殖、迁移和侵袭能力。
