State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, P.R. China.
Department of Oncology, Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang, P.R. China.
J Cell Biochem. 2018 Mar;119(3):2520-2534. doi: 10.1002/jcb.26413. Epub 2017 Nov 8.
In this study, we aim to determine the function of miR-124 on gastric cancer (GC) cells and the underlying mechanism that involves jaddeg1 (JAG1) and the Notch signaling pathway. GC tissues and adjacent tissues from 100 patients suffering from GC were selected. GC SGC-7901 and AGS cells were selected and grouped into control, mimic-NC, miR-124 mimic, inhibitor-NC, miR-124 inhibitor, and miR-124 inhibitor + si-JAG1 groups. RT-qPCR and a Western blotting assay were conducted to detect the expression of miR-124, JAG1, and Notch signaling pathway-related proteins (NICD, HES1, and HES5). MTS, wound-healing, transwell assay and flow cytometry were performed to detect cell proliferation, migration, invasion, cell cycle distribution, and apoptosis, respectively. Compared with adjacent tissues, a lower miR-124 expression and higher JAG1 expression were found in GC tissues. JAG1 is a direct target gene of miR-124. Compared with the control group, the expression of JAG1, NICD, HES1, and HES5, cell invasion, migration, and proliferation in the miR-124 mimic group were decreased, while the apoptosis rate was increased and cells were arrested at the G0/G1 phase. Compared with the miR-124 inhibitor group, the expression of JAG1, NICD, HES1, and HES5, cell invasion, migration, and proliferation in the miR-124 inhibitor + si-JAG1 group were decreased, while the apoptosis rate and cell ratio at the G0/G1 phase were increased. The demonstration that miR-124 inhibits GC cell growth supports the concept that miR-124 functions as a tumor suppressor by a mechanism that involves translational repression of the JAG1 and the inhibition of Notch signaling pathway.
在这项研究中，我们旨在确定 miR-124 在胃癌（GC）细胞中的功能以及涉及 jaddeg1（JAG1）和 Notch 信号通路的潜在机制。选择了 100 名 GC 患者的 GC 组织和相邻组织。选择 GC SGC-7901 和 AGS 细胞，并将其分为对照组、 mimic-NC 组、miR-124 模拟物组、inhibitor-NC 组、miR-124 抑制剂组和 miR-124 抑制剂+si-JAG1 组。通过 RT-qPCR 和 Western blot 检测 miR-124、JAG1 和 Notch 信号通路相关蛋白（NICD、HES1 和 HES5）的表达。通过 MTS、划痕愈合、transwell 分析和流式细胞术分别检测细胞增殖、迁移、侵袭、细胞周期分布和细胞凋亡。与相邻组织相比，GC 组织中 miR-124 表达降低，JAG1 表达升高。JAG1 是 miR-124 的直接靶基因。与对照组相比，miR-124 模拟物组中 JAG1、NICD、HES1 和 HES5 的表达、细胞侵袭、迁移和增殖减少，而细胞凋亡率增加，细胞停滞在 G0/G1 期。与 miR-124 抑制剂组相比，miR-124 抑制剂+si-JAG1 组中 JAG1、NICD、HES1 和 HES5 的表达、细胞侵袭、迁移和增殖减少，而细胞凋亡率和 G0/G1 期细胞比例增加。miR-124 抑制 GC 细胞生长的结果支持 miR-124 通过抑制翻译抑制 JAG1 和 Notch 信号通路来发挥肿瘤抑制作用的概念。
