长链非编码RNA SNHG20沉默通过从MBD1中吸附miR-5095来抑制肝细胞癌进展。
LncRNA SNHG20 silencing inhibits hepatocellular carcinoma progression by sponging miR-5095 from MBD1.
作者信息
Xu Bin, Li Chao, Yang Bin, Zhou Fan, Tang Kezhong, Wang Lantian, Lu Wenjie
机构信息
Department of Hepatopancreatobiliary Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine Hangzhou 310000, Zhejiang, China.
出版信息
Am J Transl Res. 2023 Jun 15;15(6):4314-4331. eCollection 2023.
OBJECTIVE
Long non-coding RNAs (lncRNAs) may have a significant regulatory effect on the progression of hepatocellular carcinoma (HCC), according to recent data. This study aims to investigate how SNHG20, a small nucleolar RNA host gene, contributes to the development of HCC.
METHODS
LncRNA SNHG20, miR-5095, and MBD1 gene levels were determined using reverse transcription qPCR (RT-qPCR). Huh-7 and HepG2 cell bioactivities were evaluated using the CCK-8 kit, EdU, flow cytometry, and wound-healing migration tests. To assess the metastasis of Huh-7 and HepG2 cells, a transwell assay was used. The amounts of invasion- and proliferation-associated proteins were determined using western blot. Using the miRDB (www.mirdb.org) software, the possible target genes of lncRNA and miRNA were predicted, and this prediction was then verified by a twofold luciferase reporter test. To determine the pathologic alteration and Ki67 level in tumor tissues, H&E staining and IHC were employed. TUNEL was conducted to assess the presence of apoptotic bodies in the tumor tissues.
RESULTS
lncRNA SNHG20 exhibited a high expression in HCC cells (P<0.01). LncRNA SNHG20 knockdown inhibited HCC cell metastasis (P<0.01) and accelerated apoptosis (P<0.01). LncRNA SNHG20 acted as a sponge of miR-5095 in HCC. In addition, miR-5095 overexpression inhibited HCC cell metastasis (P<0.01) and accelerated apoptosis (P<0.01); and miR-5095 negatively targeted MBD1. Furthermore, LncRNA SNHG20 regulated HCC progression through the miR-5095/MBD1 axis, and LncRNA SNHG20 knockdown inhibited HCC growth.
CONCLUSION
LncRNA SNHG20 accelerates HCC progression by the miR-5095/MBD1 axis, indicating lncRNA SNHG20 can be used as a biomarker for patients with HCC.
目的
根据近期数据，长链非编码RNA（lncRNA）可能对肝细胞癌（HCC）的进展具有显著调节作用。本研究旨在探究小核仁RNA宿主基因SNHG20如何促进HCC的发生发展。
方法
采用逆转录qPCR（RT-qPCR）检测lncRNA SNHG20、miR-5095和MBD1基因水平。使用CCK-8试剂盒、EdU、流式细胞术和伤口愈合迁移试验评估Huh-7和HepG2细胞的生物活性。采用Transwell试验评估Huh-7和HepG2细胞的转移情况。使用蛋白质免疫印迹法测定侵袭和增殖相关蛋白的含量。使用miRDB（www.mirdb.org）软件预测lncRNA和miRNA的潜在靶基因，然后通过双荧光素酶报告基因试验进行验证。采用苏木精-伊红（H&E）染色和免疫组织化学（IHC）检测肿瘤组织的病理改变和Ki67水平。采用TUNEL法评估肿瘤组织中凋亡小体的存在情况。
结果
lncRNA SNHG20在HCC细胞中高表达（P<0.01）。敲低lncRNA SNHG20可抑制HCC细胞转移（P<0.01）并加速细胞凋亡（P<0.01）。在HCC中，lncRNA SNHG20作为miR-5095的海绵发挥作用。此外，miR-5095过表达可抑制HCC细胞转移（P<0.01）并加速细胞凋亡（P<0.01）；且miR-5095负向靶向MBD1。此外，lncRNA SNHG20通过miR-5095/MBD1轴调节HCC进展，敲低lncRNA SNHG20可抑制HCC生长。
结论
lncRNA SNHG20通过miR-5095/MBD1轴加速HCC进展，表明lncRNA SNHG20可作为HCC患者的生物标志物。
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