Ai Jing, Sun Junhui, Zhou Guanhui, Zhu Tongyin, Jing Li
Department of Ophthalmology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P.R. China.
Hepatobiliary and Pancreatic Interventional Treatment Center, Division of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, P.R. China.
Cell Cycle. 2020 Apr;19(7):742-757. doi: 10.1080/15384101.2020.1729323. Epub 2020 Feb 23.
Long non-coding RNA termed GAS6 antisense RNA 1 (GAS6-AS1) plays an essential role in gastric and non-small cell lung cancers. Nonetheless, the function of GAS6-AS1 in hepatocellular carcinoma (HCC) has not been so far studied in detail. In this study, reverse-transcription quantitative PCR was performed to measure GAS6-AS1 expression in HCC samples. A series of functional experiments, including MTT assay, colony formation assay, flow-cytometric analysis, and transwell migration and invasion assays, was performed to determine the influence of GAS6-AS1 knockdown on the malignant phenotype of HCC. The results showed that GAS6-AS1 was significantly upregulated in HCC tissue samples and cell lines. Increased GAS6-AS1 expression was associated with tumor size, Edmondson grade, and Tumor-Node-Metastasis (TNM) stage among patients with HCC. The overall survival of patients with HCC characterized with high expression of GAS6-AS1 was significantly shorter in comparison to that of patients with low level of GAS6-AS1. Functional experiments indicated that knockdown of GAS6-AS1 suppressed HCC cell proliferation, colony formation, migration, and invasion ; promoted apoptosis ; and decreased tumor growth . Of note, GAS6-AS1 was validated as a competing endogenous RNA (ceRNA) for microRNA-585 (miR-585) and consequently increased the expression of eukaryotic translation initiation factor 5A2 (). Finally, rescue experiments confirmed the association among GAS6-AS1, miR-585, and EIF5A2 in HCC cells. Our study provides substantial evidence that the GAS6-AS1/miR-585/EIF5A2 pathway plays an important role in HCC progression and that might be considered as a potential target for therapeutic approaches in HCC.
名为GAS6反义RNA 1（GAS6-AS1）的长链非编码RNA在胃癌和非小细胞肺癌中发挥着重要作用。尽管如此，GAS6-AS1在肝细胞癌（HCC）中的功能迄今尚未得到详细研究。在本研究中，进行了逆转录定量PCR以检测HCC样本中GAS6-AS1的表达。进行了一系列功能实验，包括MTT法、集落形成实验、流式细胞术分析以及Transwell迁移和侵袭实验，以确定GAS6-AS1敲低对HCC恶性表型的影响。结果显示，GAS6-AS1在HCC组织样本和细胞系中显著上调。GAS6-AS1表达增加与HCC患者的肿瘤大小、Edmondson分级和肿瘤-淋巴结-转移（TNM）分期相关。与GAS6-AS1低表达的患者相比，GAS6-AS1高表达的HCC患者的总生存期显著缩短。功能实验表明，敲低GAS6-AS1可抑制HCC细胞增殖、集落形成、迁移和侵袭；促进细胞凋亡；并减少肿瘤生长。值得注意的是，GAS6-AS1被验证为微小RNA-585（miR-585）的竞争性内源RNA（ceRNA），因此增加了真核翻译起始因子5A2（）的表达。最后，挽救实验证实了HCC细胞中GAS6-AS1、miR-585和EIF5A2之间的关联。我们的研究提供了大量证据表明GAS6-AS1/miR-585/EIF5A2通路在HCC进展中起重要作用，并且可能被视为HCC治疗方法的潜在靶点。
