Wan Tao, Zheng Jun, Yao Rucheng, Yang Shuang, Zheng Weihong, Zhou Pei
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.
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.
Ann Hepatol. 2021 Jan-Feb;20:100258. doi: 10.1016/j.aohep.2020.09.003. Epub 2020 Sep 19.
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.
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.
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.
Our data revealed that DDX11-AS1 could act as an oncogenic regulator in HCC, providing a potential therapeutic target for HCC treatment.
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。
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。
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。
我们的数据表明，DDX11-AS1可能作为HCC中的致癌调节因子，为HCC治疗提供了潜在的治疗靶点。
