ABSTRACT

Background: Gastric carcinoma (GC) is a common malignant tumor. Recently, it has been found that long non-coding RNAs (lncRNAs) play important role in cancer. In this paper, we investigated the effects and mechanism of lncRNA GASL1 in GC cells.

Methods: GASL1 level in GC cells was up-regulated via cell transfection. Cell proliferation, migration, invasion were detected by CCK-8, BrdU, Transwell assays and western blot. In addition, the regulation of GASL1 on microRNA (miR)-106a level was detected using RT-qPCR and the binding between GASL1 and miR-106a was confirmed by bioinformatic prediction and luciferase reporter assay. The effects of overexpressing miR-106a on GASL1-regulated GC cell behaviors were further explored. Moreover, western blot also was used to detect the pathway-related proteins.

Results: Overexpression of GASL1 decreased the viability and BrdU levels. Meanwhile, CyclinD1 level was decreased while p53 and p21 levels were strengthened by overexpression of GASL1. On cell metastasis, up-regulation of GASL1 decreased cell migration, invasion and related proteins matrix metalloproteinase (MMP)-9 and Vimentin levels. Meanwhile, silencing GASL1 exerted opposite effects on GC cells. Moreover, GASL1 negatively regulated and targeted miR-106a. Up-regulation of miR-106a weakened the functions of GASL1 in cell proliferation and metastasis. Besides, GASL1 decreased the relate-protein levels of PI3K/AKT and ras/raf/MEK/ERK pathways while miR-106a weakened these changes.

ConclusionGASL1 restrained GC cell proliferation and metastasis and blocked PI3K/AKT and ras/raf/MEK/ERK pathways by sponging miR-106a.

摘要

背景：胃癌（GC）是一种常见的恶性肿瘤。最近，已经发现长链非编码 RNA (lncRNAs) 在癌症中起重要作用。在本文中，我们研究了 lncRNA GASL1 在 GC 细胞中的作用和机制。

方法： GC细胞中GASL1水平通过细胞转染上调。通过CCK-8、BrdU、Transwell测定和蛋白质印迹检测细胞增殖、迁移、侵袭。此外，使用 RT-qPCR 检测 GASL1 对 microRNA (miR)-106a 水平的调节，并通过生物信息学预测和荧光素酶报告基因测定证实 GASL1 和 miR-106a 之间的结合。进一步探讨了过表达 miR-106a 对 GASL1 调节的 GC 细胞行为的影响。此外，蛋白质印迹也被用于检测通路相关蛋白。

结果： GASL1 的过表达降低了生存力和 BrdU 水平。同时，过表达 GASL1 会降低 CyclinD1 水平，而增强 p53 和 p21 水平。在细胞转移中，GASL1 的上调降低了细胞迁移、侵袭和相关蛋白基质金属蛋白酶 (MMP)-9 和波形蛋白的水平。同时，沉默 GASL1 对 GC 细胞产生相反的影响。此外，GASL1 负调控和靶向 miR-106a。miR-106a 的上调减弱了 GASL1 在细胞增殖和转移中的功能。此外，GASL1 降低了 PI3K/AKT 和 ras/raf/MEK/ERK 通路的相关蛋白水平，而 miR-106a 减弱了这些变化。

结论GASL1通过海绵miR-106a抑制GC细胞增殖和转移，阻断PI3K/AKT和ras/raf/MEK/ERK通路。