长链非编码RNA DDX11-AS1通过miR-195-5p/MACC1途径加速肝癌进展。

LncRNA DDX11-AS1 accelerates hepatocellular carcinoma progression via the miR-195-5p/MACC1 pathway.

作者信息

Wan Tao, Zheng Jun, Yao Rucheng, Yang Shuang, Zheng Weihong, Zhou Pei

机构信息

The First College of Clinical Medical Science of China Three Gorges University, Yichang, Hubei, China; Institute of Hepatopancreatobilary Surgery of China Three Gorges University, Yichang, Hubei, China; Department of Hepatopancreatobilary Surgery, Yichang Central People's Hospital, Yichang, Hubei, China.

出版信息

Ann Hepatol. 2021 Jan-Feb;20:100258. doi: 10.1016/j.aohep.2020.09.003. Epub 2020 Sep 19.

DOI:10.1016/j.aohep.2020.09.003

PMID:32961346

Abstract

INTRODUCTION AND AIM

Long non-coding RNA (lncRNA) has been shown to be a vital regulator of cancer progression, including hepatocellular carcinoma (HCC). However, the role of DEAD/H box protein 11 antisense RNA 1 (DDX11-AS1) in HCC remains to be further studied.

MATERIAL AND METHODS

The expression levels of DDX11-AS1, miR-195-5p and metastasis-associated in colon cancer-1 (MACC1) were determined by quantitative real-time PCR (qRT-PCR). Cell counting kit-8 (CCK-8), transwell and apoptosis determination assays were used to evaluate cell proliferation, migration, invasion and apoptosis, respectively. Mice xenograft models were constructed to verify the effect of DDX11-AS1 on HCC tumor growth in vivo. Furthermore, lactate production, glucose consumption, ATP level and glucose uptake were detected to assess cell glucose metabolism. The interactions among DDX11-AS1, miR-195-5p and MACC1 were verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Moreover, western blot (WB) analysis was performed to evaluate the protein levels.

RESULTS

DDX11-AS1 was upregulated in HCC tissues and cells, and its silencing could inhibit HCC cell proliferation, migration, invasion and glucose metabolism, and promote apoptosis in vitro. Also, DDX11-AS1 knockdown reduced HCC tumor growth in vivo. Besides, DDX11-AS1 could interact with miR-195-5p, and miR-195-5p inhibitor reversed the inhibitory effect of silenced DDX11-AS1 on HCC cell progression. In addition, MACC1 was a target of miR-195-5p, and its overexpression reversed the suppression effect of miR-195-5p on HCC cell progression.

CONCLUSION

Our data revealed that DDX11-AS1 could act as an oncogenic regulator in HCC, providing a potential therapeutic target for HCC treatment.

摘要

引言与目的

长链非编码RNA（lncRNA）已被证明是癌症进展的重要调节因子，包括肝细胞癌（HCC）。然而，DEAD/H盒蛋白11反义RNA 1（DDX11-AS1）在HCC中的作用仍有待进一步研究。

材料与方法

通过定量实时PCR（qRT-PCR）检测DDX11-AS1、miR-195-5p和结肠癌转移相关蛋白1（MACC1）的表达水平。分别采用细胞计数试剂盒-8（CCK-8）、Transwell和凋亡检测试验评估细胞增殖、迁移、侵袭和凋亡。构建小鼠异种移植模型以验证DDX11-AS1对体内HCC肿瘤生长的影响。此外，检测乳酸生成、葡萄糖消耗、ATP水平和葡萄糖摄取以评估细胞葡萄糖代谢。通过双荧光素酶报告基因检测和RNA免疫沉淀（RIP）试验验证DDX11-AS1、miR-195-5p和MACC1之间的相互作用。此外，进行蛋白质印迹（WB）分析以评估蛋白质水平。

结果

DDX11-AS1在HCC组织和细胞中上调，其沉默可抑制HCC细胞增殖、迁移、侵袭和葡萄糖代谢，并在体外促进凋亡。此外，DDX11-AS1敲低可降低体内HCC肿瘤生长。此外，DDX11-AS1可与miR-195-5p相互作用，miR-195-5p抑制剂可逆转沉默DDX11-AS1对HCC细胞进展的抑制作用。此外，MACC1是miR-195-5p的靶标，其过表达可逆转miR-195-5p对HCC细胞进展的抑制作用。