Long noncoding RNA HAND2-AS1 reduced the viability of hepatocellular carcinoma via targeting microRNA-300/SOCS5 axis
Long noncoding RNA HAND2-AS1 reduced the viability of hepatocellular carcinoma via targeting microRNA-300/SOCS5 axis
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DOI：
10.1016/j.hbpd.2020.02.011
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发表时间：
2020-12-01
影响因子：
3.3
通讯作者：
Zhang, Hui
中科院分区：
文献类型：
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作者：
Bi, Hua-Qiang;Li, Zhong-Hui;Zhang, Hui
背景：肝细胞癌（HCC）是最常见的人类癌症之一，死亡率高。长链非编码RNA心脏和神经嵴衍生物表达2反义1 （HAND2-AS1）在包括HCC在内的多种癌症中下调，但HAND2-AS1调控HCC细胞存活的确切机制尚不清楚。方法：采用实时荧光定量PCR检测HAND2-AS1、miR-300的表达水平。Western blot检测细胞因子信号传导抑制因子5 （SOCS5）、Bcl-2、Bax和cleaved caspase-3蛋白水平。分别用细胞计数试剂盒-8和克隆形成法测定细胞活力和细胞增殖。流式细胞术检测细胞凋亡。HAND2-AS1与miR-300、miR-300与SOCS5之间的相互作用通过荧光素酶报告基因试验验证。结果：HAND2-AS1在HCC组织和细胞系中表达下调，且其表达水平与患者生存率呈正相关。HAND2-AS1过表达降低HCC细胞的活力和增殖。HAND2-AS1水平升高可诱导HCC细胞凋亡，同时Bax和cleaved caspase-3水平升高，Bcl-2水平降低。我们还验证了HAND2-AS1作为miR-300的海绵，并且在HCC组织中HAND2-AS1与miR-300的表达水平呈负相关。此外，我们发现SOCS5是miR-300的下游靶点。此外，miR-300模拟物消除了hand2 - as1介导的细胞活力和增殖抑制。miR-300模拟物也逆转了hand2 - as1诱导的HCC细胞凋亡。结论：lncRNA HAND2-AS1通过调控miR-300/SOCS5轴抑制HCC细胞增殖。(C) 2020浙江大学医学院第一附属医院Elsevier B.V.版权所有。
Background: Hepatocellular carcinoma (HCC) is one of the most prevalent human cancers with high mortality. Long non-coding RNA heart and neural crest derivatives expressed 2 anti-sense 1 (HAND2-AS1) is down-regulated in several cancers including HCC, yet the precise mechanisms how HAND2-AS1 regulates cell survival in HCC remains poorly understood.Methods: The expression levels of HAND2-AS1 and miR-300 were measured using quantitative real-time PCR. The protein levels of suppressor of cytokine signaling 5 (SOCS5), Bcl-2, Bax and cleaved caspase-3 were determined by Western blot. Cell viability and cell proliferation were assessed using cell counting kit-8 and clone formation assay, respectively. Cell apoptosis was detected using flow cytometry. The interactions between HAND2-AS1 and miR-300, miR-300 and SOCS5 were validated using luciferase reporter assay.Results: HAND2-AS1 was down-regulated in HCC tissues and cell lines, and the expression level of HAND2-AS1 was positively correlated to patient survival. HAND2-AS1 over-expression reduced viability and proliferation in HCC cells. Elevated HAND2-AS1 level induced apoptosis in HCC cells, accompanied with increased Bax and cleaved caspase-3 levels and decreased Bcl-2 level. We also validated that HAND2-AS1 acted as a sponge of miR-300, and there was a negative correlation between expression levels of HAND2-AS1 and miR-300 in HCC tissues. Furthermore, we found that SOCS5 was a downstream target of miR-300. In addition, miR-300 mimics abolished HAND2-AS1-mediated inhibition of cell viability and proliferation. miR-300 mimics also reversed the HAND2-AS1-induced apoptosis in HCC cells.Conclusion: lncRNA HAND2-AS1 inhibits proliferation in HCC through regulating miR-300/SOCS5 axis. (C) 2020 First Affiliated Hospital, Zhejiang University School of Medicine in China. Published by Elsevier B.V. All rights reserved.