Effects of hsa_circ_0000711 expression level on proliferation and apoptosis of hepatoma cells
Effects of hsa_circ_0000711 expression level on proliferation and apoptosis of hepatoma cells
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DOI：
10.26355/eurrev_202004_20996
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发表时间：
2020-04-01
影响因子：
3.3
通讯作者：
Huang, J. -N.
中科院分区：
文献类型：
--
作者：
Chen, K. -H.;Pan, J. -F.;Huang, J. -N.
目的：探讨人血清白蛋白（hsa）_circular（circ）_0000711在肝细胞癌（hepatocellular carcinoma，HCC）中的作用.环形核糖核酸（Circular ribonucleic acids，circRNA）在肿瘤生物学中发挥着重要作用，但其在HCC中的作用在很大程度上仍不清楚。患者与方法：应用circRNA表达谱芯片筛选HCC患者癌组织及癌旁组织中差异表达的circRNA，并采用逆转录-定量聚合酶链反应（RT-qPCR）进行进一步验证。然后，利用生物信息学软件预测关键circRNA的靶microRNA（miRNAs）及其信使RNA（mRNAs），构建circRNA-miRNAs-mRNA调控网络。随后，应用KEGG和GO富集分析来预测hsa_circ_0000711及其相关信号通路可能调控的生物学过程。然后选择在circRNA-miR-NA-mRNA调控网络中起关键作用的miRNAs作为研究对象。并通过荧光素酶报告基因测定证实它们与HSA_Circ_0000711的直接结合。此后，hsa_circ_0000711被过表达或敲除。CCK-8法检测hsa_circ_0000711的生物学功能。细胞凋亡检测和体外5-乙炔基-2 '-脱氧尿苷（EdU）染色检测。结果：表达谱筛选结果显示，肝癌患者肿瘤组织和癌旁组织中circRNA的表达谱存在显着差异。基于circRNA表达谱和RT-qPCR结果，hsa_circ_0000711的表达水平在HCC组织中明显降低。另外。miR-103 a-3 p在circRNA-miRNA-mRNA调控网络中具有最高的特征向量中心性。提示miR-103 a-3 p是hsa_circ_0000711病理机制的重要参与者。KEGG富集分析结果指出，hsa_circ_0000711调控的靶基因在肿瘤相关信号通路中明显富集。此外，GO富集分析的结果表明，hsa_circ_0000711调节的生物学过程主要与细胞周期调节有关，因此可能会影响细胞增殖。荧光素酶报告基因和RT-qPCR检测结果显示hsa_circ_0000711直接与has-miR-103 a-3 p结合，起到分子海绵的作用。CCK-8和EdU染色结果显示hsa_circ_0000711过表达组肝癌细胞增殖明显增强。结论：hsa_circ_0000711过表达可通过靶向has-miR-103 a-3 p促进肝癌细胞增殖，抑制肝癌细胞凋亡。
OBJECTIVE: To investigate the role of human serum albumin (hsa)_circular (circ)_0000711 in hepatocellular carcinoma (HCC). Circular ribonucleic acids (circRNAs) are proven in numerous studies to play crucial role in tumor biology, but their roles in HCC remain unknown to a great extent.PATIENTS AND METHODS: The circRNA expression profile microarray was employed to screen differentially expressed circRNAs in tumor tissues and adjacent tissues from HCC patients, and Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) assay was performed for further verification. Next, the target micro RNAs (miRNAs) and their messenger RNAs (mRNAs) of key circRNAs were predicted by bioinformatics software, and a circRNA-miRNA-mRNA regulatory network was constructed. Subsequently, KEGG and GO enrichment analyses were applied to predict the possible biological processes regulated by hsa_circ_0000711 and relevant signaling pathways. The miRNAs playing a key role in the circRNA-miR-NA-mRNA regulatory network were then selected as the objects. and their direct binding to hsa_circ_0000711 was confirmed via luciferase reporter gene assay. Thereafter, hsa_circ_0000711 was overexpressed or knocked out. and the biological function of hsa_circ_0000711 was detected by cell counting kit-8 (CCK-8) assay. apoptosis detection, and 5-Ethynyl-2'-deoxyuridine (EdU) staining assay in vitro.RESULTS: The results of expression profile screening revealed that there was a significant difference in the expression profile of circRNAs between tumor tissues and adjacent tissues in HCC patients. Based on the circRNA expression profile and RT-qPCR results, the expression level of hsa_circ_0000711 was overtly reduced in HCC tissues. In addition. miR-103a-3p had the highest eigenvector centrality in the circRNA-miRNA-mRNA regulatory network. suggesting that miR-103a-3p is a vital participant in the pathological mechanism of hsa_circ_0000711. The KEGG enrichment analysis results pointed out that the target genes regulated by hsa_circ_0000711 were clearly enriched in the tumor-associated signaling pathways. Besides, the results of GO enrichment analysis demonstrated that the biological processes regulated by hsa_circ_0000711 were mainly related to cell cycle regulation, so cell proliferation might be affected. The results of luciferase reporter gene and RT-qPCR assays showed that hsa_circ_0000711 directly bound to has-miR-103a-3p to serve as a molecular sponge. The results of CCK-8 and EdU staining assays revealed that the proliferation of hepatoma cells in hsa_circ_0000711 overexpression group was evidently enhanced. In addition, it was further found via flow cytometry that the apoptosis rate of cells was significantly raised in hsa_circ_0000711 low-expression group and dramatically declined in hsa_circ _0000711 overexpression group.CONCLUSIONS: Overexpression of hsa_circ_0000711 promoted the proliferation and inhibited the apoptosis of hepatoma cells via targeting has-miR-103a-3p.