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环状 RNA 0091579 通过 miR-1270/YAP1 轴抑制 HCC 增殖和谷氨酰胺代谢。

Circ_0091579 Knockdown Inhibited HCC Proliferation and Glutamine Metabolism Through miR-1270/YAP1 Axis.

机构信息

Department of Radiotherapy, Taian City Central Hospital, No. 29, Longtan Road, Taishan District, Tai'an, Shandong, China.

出版信息

Biochem Genet. 2024 Feb;62(1):208-228. doi: 10.1007/s10528-023-10386-w. Epub 2023 Jun 14.

Abstract

A growing number of studies have indicated that circRNAs play an important role in the progression of malignant tumors, including hepatocellular carcinoma (HCC). In this study, we designed to explore the abnormal expression of hsa_circ_0091579 (circ_0091579) and its role in the pathogenesis of HCC. In this study, the mRNA levels of circ_0091579, miR-1270, and Yes-associated protein (YAP1) were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). RNase R and Actinomycin D were used to test the stability of circ_0091579. Cell Counting Kit-8 (CCK-8) was used to measure cell viability. Tubule formation assay was used to determine the effect of HCC cells on the number of tubes. Cell apoptosis was detected by flow cytometry. Western blot was used for the protein levels. Transwell and wound healing tests were used to measure the abilities of invasion and migration. The effect of circ_0091579 knockdown on tumor growth was verified in vivo by xenograft tumor assay and Immunohistochemistry (IHC) analysis. Dual-luciferase reporter or RIP assay was used to detect the relationship between miR-1270 and circ_0091579 or YAP1. Glutamine metabolism was determined by ELISA and western blot assays. In the present study, we found that circ_0091579 was upregulated in HCC tissues and cells. Inhibited circ_0091579 expression significantly suppressed proliferation and promoted apoptosis of HCC cells. Moreover, circ_0091579 knockdown inhibited tumor growth in vivo. Bioinformatic prediction and luciferase assay showed that circ_0091579 acted as a molecular sponge for miR-1270 and YAP1 was a target gene of miR-1270. MiR-1270 silencing could reverse the inhibitory effect of circ_0091579 knockdown on HCC progression, and YAP1 overexpression also could reverse the suppressive effect of circ_0091579 silencing on HCC progression. Meanwhile, miR-1270 inhibitor could invert the negative regulation effect of circ_0091579 silencing on YAP1 expression. Circ_0091579 promoted HCC progression by regulating the miR-1270/YAP1 axis, and our study might offer novel biomarkers and therapeutic targets for HCC.

摘要

越来越多的研究表明 circRNAs 在恶性肿瘤的进展中发挥着重要作用,包括肝细胞癌(HCC)。在这项研究中,我们旨在探讨 hsa_circ_0091579(circ_0091579)的异常表达及其在 HCC 发病机制中的作用。在这项研究中,通过实时定量聚合酶链反应(qRT-PCR)评估 circ_0091579、miR-1270 和 Yes 相关蛋白(YAP1)的 mRNA 水平。使用核糖核酸酶 R 和放线菌素 D 来测试 circ_0091579 的稳定性。使用细胞计数试剂盒-8(CCK-8)测量细胞活力。管形成测定用于确定 HCC 细胞对管数的影响。通过流式细胞术检测细胞凋亡。使用蛋白质印迹法检测蛋白质水平。Transwell 和伤口愈合试验用于测量侵袭和迁移能力。通过异种移植肿瘤测定和免疫组织化学(IHC)分析在体内验证 circ_0091579 敲低对肿瘤生长的影响。双荧光素酶报告或 RIP 测定用于检测 miR-1270 与 circ_0091579 或 YAP1 之间的关系。通过 ELISA 和 Western blot 测定测定谷氨酰胺代谢。在本研究中,我们发现 circ_0091579 在 HCC 组织和细胞中上调。抑制 circ_0091579 的表达显着抑制 HCC 细胞的增殖并促进其凋亡。此外,circ_0091579 敲低抑制体内肿瘤生长。生物信息学预测和荧光素酶测定表明 circ_0091579 作为 miR-1270 的分子海绵,而 YAP1 是 miR-1270 的靶基因。沉默 miR-1270 可以逆转 circ_0091579 敲低对 HCC 进展的抑制作用,而过表达 YAP1 也可以逆转 circ_0091579 沉默对 HCC 进展的抑制作用。同时,miR-1270 抑制剂可以反转 circ_0091579 沉默对 YAP1 表达的负调节作用。Circ_0091579 通过调节 miR-1270/YAP1 轴促进 HCC 进展,我们的研究可能为 HCC 提供新的生物标志物和治疗靶点。

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