长链非编码 RNA DCST1-AS1 通过 AKT/mTOR 信号通路加速肝癌细胞的增殖、转移和自噬。

LncRNA DCST1-AS1 accelerates the proliferation, metastasis and autophagy of hepatocellular carcinoma cell by AKT/mTOR signaling pathways.

机构信息

Department of Hepatobiliary Surgery, Affiliated Min da Hospital of Hubei Institute for Nationalities, Enshi, Hubei Province, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Jul;23(14):6091-6104. doi: 10.26355/eurrev_201907_18423.

DOI:10.26355/eurrev_201907_18423

PMID:31364110

Abstract

OBJECTIVE

The previous research revealed that long noncoding RNAs (lncRNAs) play a vital role in the development of hepatocellular carcinoma (HCC). To further discuss the underlying mechanisms of lncRNA DCST1-AS1 in the pathogenesis of HCC.

PATIENTS AND METHODS

We screened the abnormally expressed genes in HCC tissues through microarray analysis and found that lncRNA DCST1-AS1 was one of the genes significantly up-regulated. Real Time-Polymerase Chain Reaction (RT-qPCR) was used to test the gene expression of lncRNA DCST1-AS1 in HCC tissues and HepG2 cells. Respectively, CCK-8 assay, flow cytometry detection, transwell assay, wound healing assay, transmission electron microscopy, and immunofluorescence staining were used to assess the proliferation, apoptosis, migration, and autophagy of HepG2 cells. Meanwhile, the expression of signaling pathway proteins was detected by Western blot.

RESULTS

LncRNA DCST1-AS1 was confirmed hyper-expression both in HCC tissues and HCC cells. High expression of lncRNA DCST1-AS1 was significantly correlated with inferior prognosis. Moreover, lncRNA DCST1-AS1 depletion suppressed proliferation and accelerated apoptosis, activated cycle arrest, restrained cell migration, and stimulated autophagy in HCC cells. In addition, it is found that the depletion of lncRNA DCST1-AS1 on HepG2 cells exhibits anti-tumor characteristics and was mediated by the AKT/mTOR signal transduction pathway. Furthermore, pre-treated HepG2 cells with SC79, an AKT signal activator, partially restored the effect of lncRNA DCST1-AS1 silencing on HepG2 cell proliferation, apoptosis, migration, and autophagy.

CONCLUSIONS

Our results suggested that lncRNA DCST1-AS1, as a carcinogenic factor in HCC, promoted cell proliferation, and invasion, inhibited apoptosis and autophagy by modulating the AKT/mTOR signaling cascade. Therefore, our findings showed that lncRNA DCST1-AS1 may improve potential treatment strategies for HCC.

摘要

目的

先前的研究表明，长非编码 RNA（lncRNA）在肝细胞癌（HCC）的发展中发挥着重要作用。为了进一步探讨 lncRNA DCST1-AS1 在 HCC 发病机制中的潜在机制。

患者和方法

我们通过微阵列分析筛选 HCC 组织中异常表达的基因，发现 lncRNA DCST1-AS1 是显著上调的基因之一。实时聚合酶链反应（RT-qPCR）用于检测 HCC 组织和 HepG2 细胞中 lncRNA DCST1-AS1 的基因表达。分别采用 CCK-8 测定、流式细胞术检测、Transwell 测定、划痕愈合测定、透射电子显微镜和免疫荧光染色来评估 HepG2 细胞的增殖、凋亡、迁移和自噬。同时，通过 Western blot 检测信号通路蛋白的表达。

结果

lncRNA DCST1-AS1 在 HCC 组织和 HCC 细胞中均被证实呈高表达。lncRNA DCST1-AS1 的高表达与预后不良显著相关。此外，lncRNA DCST1-AS1 耗竭抑制 HCC 细胞的增殖并加速其凋亡，激活细胞周期停滞，抑制细胞迁移，并刺激自噬。此外，发现 HepG2 细胞中 lncRNA DCST1-AS1 的耗竭表现出抗肿瘤特性，并通过 AKT/mTOR 信号转导通路介导。此外，用 AKT 信号激活剂 SC79 预处理 HepG2 细胞可部分恢复 lncRNA DCST1-AS1 沉默对 HepG2 细胞增殖、凋亡、迁移和自噬的影响。