Circular RNA circ-102,166 acts as a sponge of miR-182 and miR-184 to suppress hepatocellular carcinoma proliferation and invasion
Circular RNA circ-102,166 acts as a sponge of miR-182 and miR-184 to suppress hepatocellular carcinoma proliferation and invasion
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环状RNA circ-102,166充当miR-182和miR-184的海绵，抑制肝细胞癌的增殖和侵袭
DOI：
10.1007/s13402-020-00564-y
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发表时间：
2021
影响因子：
6.6
通讯作者：
Liu Wei
中科院分区：
文献类型：
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作者：
Li Rong;Deng Yinan;Liang Jinliang;Hu Zhongying;Li Xuejiao;Liu Huanyi;Wang Guoying;Fu Binsheng;Zhang Tong;Zhang Qi;Yang Yang;Chen Guihua;Liu Wei
目的肝细胞癌（HCC）中存在多个环状RNA（circRNAs）的表达异常。然而，它们的功能和行动方式在很大程度上仍不清楚。方法通过整合3个人肝癌circRNA微阵列数据集（GSE 94508、GSE 97332和GSE 78520），在原发性肝癌组织和细胞系中进行qRT-PCR验证，鉴定肝癌中表达异常的circRNA。通过桑格测序、RNase R处理、北方印迹和细胞内定位分析来验证circRNA特征。此外，使用CCK 8、集落形成、EDU掺入、流式细胞术、体外transwell和划痕伤口愈合测定以及体内肿瘤异种移植测定来评估HCC发展中的circRNA功能。接下来，潜在的分子机制在肝癌进行了评估，使用双荧光素酶报告，RNA下拉，RNA免疫沉淀和蛋白质印迹assessment.ResultsWe发现，一种新的环状RNA，环102，166，下调在肝癌，其表达水平与多种临床病理特征，以及肝癌患者的临床预后显着相关。体内外实验表明，circ-102，166过表达可显著抑制肝癌细胞的增殖、侵袭、迁移和致瘤性。此外，我们发现circ-102，166可以与miR-182和miR-184结合，调节其下游靶点FOXO 3a、MTSS 1、SOX 7、p-RB和c-MYC的表达。circ-102，166的下调通过释放oncomiRs miR-182和miR-184增强HCC细胞的增殖和侵袭。
PurposeMultiple circular RNAs (circRNAs) have been reported to be dysregulated in hepatocellular carcinoma (HCC). However, their functions and modes of action are still largely unclear. Identifying key circRNAs and revealing their potential functions and molecular mechanisms is considered important for improving the diagnosis and treatment of HCC.MethodsDysregulated circRNAs in HCC were identified through integration of three human HCC circRNAs microarray datasets (GSE94508, GSE97332 and GSE 78520), followed by qRT-PCR validation in primary HCC tissues and cell lines. circRNA characteristics were verified through Sanger sequencing, RNase R treatment, northern blotting and intracellular localization analyses. In addition, circRNA functions in HCC development were assessed using CCK8, colony formation, EDU incorporation, flow cytometry, transwell and scratch wound healing assays in vitro and tumor xenograft assays in vivo. Next, underlying molecular mechanisms in HCC were assessed using dual-luciferase reporter, RNA pull-down, RNA immunoprecipitation and western blotting assays.ResultsWe found that a novel circular RNA, circ-102,166, was down-regulated in HCC and that its expression level was significantly associated with multiple clinicopathologic characteristics, as well as the clinical prognosis of HCC patients. In vitro and in vivo experiments revealed that circ-102,166 overexpression significantly inhibited the proliferation, invasion, migration and tumorigenicity of HCC cells. Furthermore, we found that circ-102,166 can bind to miR-182 and miR-184 to regulate the expression of several of their downstream targets (FOXO3a, MTSS1, SOX7, p-RB and c-MYC).ConclusionOur data revealed a tumor-suppressing role of circ-102,166 in HCC. Down-regulation of circ-102,166 enhanced the proliferation and invasion of HCC cells by releasing the oncomiRs miR-182 and miR-184.