Circular RNA CircEPB41L2 Functions as Tumor Suppressor in Hepatocellular Carcinoma Through Sponging miR-590-5p.
Circular RNA CircEPB41L2 Functions as Tumor Suppressor in Hepatocellular Carcinoma Through Sponging miR-590-5p.
复制标题
环状 RNA CircEPB41L2 通过海绵 miR-590-5p 在肝细胞癌中发挥肿瘤抑制作用
DOI：
10.2147/cmar.s291682
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发表时间：
2021
影响因子：
3.3
通讯作者：
Cai Z
中科院分区：
文献类型：
--
作者：
Chen F;He L;Qiu L;Zhou Y;Li Z;Chen G;Xin F;Dong X;Xu H;Wang G;Liu J;Cai Z
研究背景环状RNA（circular RNAs，circRNA）可与miRNA相互作用调节基因表达，参与肝细胞癌（hepatocellular carcinoma，HCC）的发生发展。本研究旨在探讨circEPB 41 L2（hsa_circ_0077837）在肝癌进展过程中的潜在功能和分子机制。材料和方法采用实时定量PCR（qRT-PCR）对肝癌组织和肝癌细胞系中circEPB 41 L2的表达进行定量。CCK-8法和集落形成法检测肝癌细胞增殖情况。通过创伤愈合实验和transwell实验检测肝癌细胞的迁移和侵袭能力。进行蛋白质印迹以确定蛋白质水平上的基因表达。通过异种移植实验研究circEPB 41 L2对体内HCC的作用。通过生物信息学方法预测circEPB 41 L2与miR-590- 5 p之间的相互作用，并通过荧光素酶报告基因测定证实。结果对61例HCC患者的肿瘤和癌旁组织中circRNA谱的广泛分析表明，circEPB 41 L2在HCC中显著下调，这在另一组HCC中通过qRT-PCR分析进一步证实。临床病理分析显示，circEPB 41 L2表达下调与肿瘤大小、血管浸润和甲胎蛋白呈负相关，与肝癌预后呈正相关。体外生物学功能实验表明，过表达circEPB 41 L2可明显抑制肝癌细胞的增殖和转移，而敲低circEPB 41 L2则相反。此外，我们还发现circEPB 41 L2通过EMT信号通路抑制HCC的迁移和侵袭。同样，circEPB 41 L2的过表达也可以显著抑制体内HCC细胞的增殖。生物信息学分析和荧光素酶报告基因分析表明，circEPB 41 L2与miR-590- 5 p直接相互作用，并在miRNA拯救实验中验证了相应的生物学功能。结论circEPB 41 L2可能通过吞噬miR-590- 5 p在肝癌进展过程中发挥抑癌作用。
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.