环状RNA CircEPB41L2通过海绵吸附miR-590-5p在肝细胞癌中发挥肿瘤抑制作用。
Circular RNA CircEPB41L2 Functions as Tumor Suppressor in Hepatocellular Carcinoma Through Sponging miR-590-5p.
作者信息
Chen Feng, He Lei, Qiu Liman, Zhou Yang, Li Zhenli, Chen Geng, Xin Fuli, Dong Xiuqing, Xu Haipo, Wang Gaoxiong, Liu Jingfeng, Cai Zhixiong
机构信息
College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China.
The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, People's Republic of China.
出版信息
Cancer Manag Res. 2021 Apr 1;13:2969-2981. doi: 10.2147/CMAR.S291682. eCollection 2021.
BACKGROUND
Circular RNAs (circRNAs) could interact with miRNAs to regulate gene expression, participating in hepatocellular carcinoma (HCC) initiation and development. This work aimed to determine the potential function and molecular mechanism of circEPB41L2 (hsa_circ_0077837) during HCC progression.
MATERIALS AND METHODS
The expression of circEPB41L2 in HCC tissues and HCC cell lines was quantified using real-time quantitative PCR (qRT-PCR). CCK-8 assays and colony formation assays were utilized to detect the proliferation of HCC cells. Wound healing assay and transwell assay were performed to determine the capability of migration and invasion for HCC cells. Western blot was conducted to determine gene expression on protein levels. The effect of circEPB41L2 on HCC in vivo was investigated via xenograft experiment. Interaction between circEPB41L2 and miR-590-5p was predicted through bioinformatics methods and confirmed via luciferase reporter assay.
RESULTS
Extensive analysis of circRNA profiles in tumor and matched para-tumor tissues collected from 61 HCC patients identified that circEPB41L2 was significantly down-regulated in HCC, which was further confirmed in another HCC group by qRT-PCR analysis. The clinicopathological analysis revealed that down-regulation of circEPB41L2 was negatively associated with tumor size, vascular invasion and alpha-fetoprotein, while positively correlated with HCC prognosis. The biological function experiments showed that overexpression of circEPB41L2 could obviously inhibit the proliferation and metastasis of HCC cells in vitro, while knockdown of circEPB41L2 induced opposite results. Moreover, we also found that circEPB41L2 inhibited HCC migration and invasion though EMT signaling pathway. Similarly, overexpression of circEPB41L2 can also significantly inhibit the proliferation of HCC cells in vivo. Bioinformatic analysis and luciferase reporter assay revealed that circEPB41L2 interacts directly with miR-590-5p and the corresponding biological functions were also verified in miRNA rescue experiments.
CONCLUSION
Our results suggest that circEPB41L2 might function as a tumor suppressor during HCC progression by sponging miR-590-5p.
背景
环状RNA（circRNAs）可与微小RNA（miRNAs）相互作用以调节基因表达，参与肝细胞癌（HCC）的发生和发展。本研究旨在确定circEPB41L2（hsa_circ_0077837）在HCC进展过程中的潜在功能和分子机制。
材料与方法
采用实时定量聚合酶链反应（qRT-PCR）定量检测circEPB41L2在HCC组织和HCC细胞系中的表达。使用CCK-8法和集落形成试验检测HCC细胞的增殖情况。进行伤口愈合试验和Transwell试验以确定HCC细胞的迁移和侵袭能力。通过蛋白质印迹法检测基因在蛋白质水平的表达。通过异种移植实验研究circEPB41L2对体内HCC的影响。通过生物信息学方法预测circEPB41L2与miR-590-5p之间的相互作用，并通过荧光素酶报告基因试验进行验证。
结果
对61例HCC患者的肿瘤组织和配对的癌旁组织进行circRNA谱的广泛分析发现，circEPB41L2在HCC中显著下调，qRT-PCR分析在另一组HCC中进一步证实了这一结果。临床病理分析显示，circEPB41L2的下调与肿瘤大小、血管侵犯和甲胎蛋白呈负相关，而与HCC预后呈正相关。生物学功能实验表明，circEPB41L2的过表达可明显抑制体外HCC细胞的增殖和转移，而circEPB41L2的敲低则产生相反的结果。此外，我们还发现circEPB41L2通过上皮-间质转化（EMT）信号通路抑制HCC的迁移和侵袭。同样，circEPB41L2的过表达也可显著抑制体内HCC细胞的增殖。生物信息学分析和荧光素酶报告基因试验表明，circEPB41L2直接与miR-590-5p相互作用，并且在miRNA拯救实验中也验证了相应的生物学功能。
结论
我们的结果表明，circEPB41L2可能通过吸附miR-590-5p在HCC进展过程中发挥肿瘤抑制作用。
Zotero 插件