Department of Oncology, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
Eur Rev Med Pharmacol Sci. 2020 Apr;24(8):4161-4171. doi: 10.26355/eurrev_202004_20996.
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.
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-miRNA-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.
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.
Overexpression of hsa_circ_0000711 promoted the proliferation and inhibited the apoptosis of hepatoma cells via targeting has-miR-103a-3p.
探讨人血清白蛋白（hsa）_环状（circ）_0000711 在肝细胞癌（HCC）中的作用。大量研究证明，环状核糖核酸（circRNAs）在肿瘤生物学中发挥着重要作用，但它们在 HCC 中的作用在很大程度上仍不清楚。
采用 circRNA 表达谱微阵列筛选 HCC 患者肿瘤组织和相邻组织中差异表达的 circRNAs，并用逆转录定量聚合酶链反应（RT-qPCR）检测进行进一步验证。接下来，利用生物信息学软件预测关键 circRNAs 的靶微小 RNA（miRNA）及其信使 RNA（mRNA），构建 circRNA-miRNA-mRNA 调控网络。随后，应用 KEGG 和 GO 富集分析预测 hsa_circ_0000711 调控的可能生物学过程及相关信号通路。然后选择在 circRNA-miRNA-mRNA 调控网络中起关键作用的 miRNAs 作为对象，并通过荧光素酶报告基因检测证实它们与 hsa_circ_0000711 的直接结合。此后，过表达或敲除 hsa_circ_0000711，通过细胞计数试剂盒-8（CCK-8）检测、凋亡检测和 5-乙炔基-2'-脱氧尿苷（EdU）染色检测，体外检测 hsa_circ_0000711 的生物学功能。
表达谱筛选结果显示，HCC 患者肿瘤组织和相邻组织的 circRNAs 表达谱存在显著差异。基于 circRNA 表达谱和 RT-qPCR 结果，hsa_circ_0000711 在 HCC 组织中表达明显下调。此外，在 circRNA-miRNA-mRNA 调控网络中，miR-103a-3p 的特征向量中心度最高，表明 miR-103a-3p 是 hsa_circ_0000711 病理机制中的重要参与者。KEGG 富集分析结果表明，hsa_circ_0000711 调控的靶基因明显富集在肿瘤相关信号通路中。此外，GO 富集分析结果表明，hsa_circ_0000711 调控的生物学过程主要与细胞周期调控有关，因此细胞增殖可能受到影响。荧光素酶报告基因和 RT-qPCR 检测结果表明，hsa_circ_0000711 可直接结合 has-miR-103a-3p 作为分子海绵。CCK-8 和 EdU 染色检测结果显示，hsa_circ_0000711 过表达组肝癌细胞的增殖明显增强。此外，流式细胞术进一步发现，hsa_circ_0000711 低表达组细胞凋亡率显著升高，hsa_circ_0000711 过表达组细胞凋亡率显著降低。
hsa_circ_0000711 通过靶向 has-miR-103a-3p 促进肝癌细胞增殖，抑制其凋亡。
