LncRNA DCST1-AS1 通过靶向 miR-605-3p 调控胃癌细胞的增殖、迁移、侵袭和凋亡。

LncRNA DCST1-AS1 regulated cell proliferation, migration, invasion and apoptosis in gastric cancer by targeting miR-605-3p.

机构信息

Department of Gastrointestinal and Colorectal Surgery, China-Japan Union Hospital of Jilin University, Economic and Technological Development Zone, Changchun City, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Feb;24(3):1158-1167. doi: 10.26355/eurrev_202002_20167.

DOI:10.26355/eurrev_202002_20167

PMID:32096164

Abstract

OBJECTIVE

Gastric cancer (GC) is one of the most common cancers in the world, with a high incidence and a poor prognosis. A large number of lncRNAs have been demonstrated to play multiple important roles in cancer development and progression. LncRNA is usually used as ceRNA and forms a regulatory network with miRNA in gastric cancer. However, the function and regulatory network of lncRNA in gastric cancer have not been fully elucidated.

MATERIALS AND METHODS

The qRT-PCR assay was used to detect DCST1-AS1 and miR-605-3p expression. Western blot was applied to measure the protein expression of CDK4, cyclin D1, MMP-2, MMP-9, cleaved caspase 3, Bcl-2, Bax and β-actin. MTT assay and flow cytometry were performed to assess cell proliferation and apoptosis, respectively. Transwell migration and invasion assay were used to determine cell migration capacity and invasion ability. Luciferase reporter assay was applied to determine the relationship of DCST-AS1 and miR-605-3p in GC.

RESULTS

In this study, we found that DCST1-AS1 was highly expressed while miR-605-3p was low expressed in GC tissues and cells. Moreover, DCST1-AS1 expression negatively regulated miR-605-3p expression in GC. Functionally test demonstrated that knockdown of DCST1 inhibited cell proliferation, migration and invasion as well as promoted cell apoptosis in GC cells. Interestingly, miR-605-3p has been verified to be a target miRNA of DCST1-AS1 with luciferase reporter assay. More than that, the reverse experiment determined that the inhibition of miR-605-3p could alleviate the suppressive effects of low DCST1-AS1 expression on cell growth in GC.

CONCLUSIONS

We proved the regulatory network of lncRNA DCST1-AS1 for the first time, and also explored and found that lncRNA DCST1-AS1 regulated cell proliferation, migration, invasion and apoptosis by regulation of miR-605-3p, providing a new therapeutic target for gastric cancer treatment.

摘要

目的

胃癌（GC）是世界上最常见的癌症之一，发病率高，预后差。大量长链非编码 RNA（lncRNA）已被证明在癌症的发生和发展中发挥着多种重要作用。lncRNA 通常作为 ceRNA 并与 miRNA 在胃癌中形成调节网络。然而，lncRNA 在胃癌中的功能和调节网络尚未完全阐明。

材料和方法

采用 qRT-PCR 检测 DCST1-AS1 和 miR-605-3p 的表达。Western blot 检测 CDK4、cyclin D1、MMP-2、MMP-9、cleaved caspase 3、Bcl-2、Bax 和 β-actin 的蛋白表达。MTT 检测和流式细胞术分别评估细胞增殖和凋亡。Transwell 迁移和侵袭实验用于确定细胞迁移能力和侵袭能力。荧光素酶报告实验用于确定 DCST-AS1 和 miR-605-3p 在 GC 中的关系。

结果

本研究发现，DCST1-AS1 在 GC 组织和细胞中高表达，而 miR-605-3p 低表达。此外，DCST1-AS1 表达负调控 GC 中 miR-605-3p 的表达。功能试验表明，敲低 DCST1 可抑制 GC 细胞的增殖、迁移和侵袭，并促进细胞凋亡。有趣的是，荧光素酶报告实验验证了 miR-605-3p 是 DCST1-AS1 的靶 miRNA。此外，逆转实验确定了抑制 miR-605-3p 可以减轻低 DCST1-AS1 表达对 GC 细胞生长的抑制作用。