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circ_0088046 通过 circ_0088046-miR-1299-RTKN2 ceRNA 通路抑制肝癌细胞生长和迁移。

Depletion of circ_0088046 suppressed cell growth and motility of hepatocellular carcinoma via circ_0088046-miR-1299-RTKN2 ceRNA pathway.

机构信息

Department of General Surgery, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, People's Republic of China.

Department of Breast Cancer, Shaanxi Provincial Cancer Hospital, Xi'an, People's Republic of China.

出版信息

J Viral Hepat. 2023 Oct;30(10):819-829. doi: 10.1111/jvh.13870. Epub 2023 Jul 8.

Abstract

Circular RNAs (circRNAs) have been verified to be important modulators and therapeutic targets of human hepatocellular carcinoma (HCC). This study aims to explore the role and mechanism of circ_0088046 in HCC progression. Quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry assays were used to detect the mRNA and protein expression of circ_0088046, miR-1299, Rhotekin 2 (RTKN2), Bax, Bcl-2, E-cadherin and Ki-67. Cell proliferation was investigated by 5-Ethynyl-2'-deoxyuridine (EdU) assay and cell colony formation assay. Cell apoptosis rate was measured by flow cytometry. Transwell migration and invasion assays were adopted to assess cell migration and invasion. The molecular target relationship between miR-1299 and circ_0088046 or RTKN2 were analysed by dual-luciferase reporter assay and RNA immunoprecipitation assay. An animal experiment was conducted to demonstrate the effect of circ_0088046 on tumour formation in vivo. High levels of circ_0088046 and RTKN2, and low levels of miR-1299 were displayed in HCC tissues and cells. Circ_0088046 absence repressed cell proliferation, migration and invasion, but boosted apoptosis of HCC cells. MiR-1299 was a target of circ_0088046 and miR-1299 inhibitor restored circ_0088046 silencing-mediated inhibitory impacts on HCC cell malignancy. MiR-1299 could directly target RTKN2, and overexpressed RTKN2 rescued the suppressive effects caused by miR-1299 mimic. In addition, circ_0088046 silencing constrained tumour formation in vivo. Circ_0088046 contributed to HCC cell malignancy via modulating the miR-1299/RTKN2 axis.

摘要

环状 RNA(circRNAs)已被证实是人类肝细胞癌(HCC)的重要调节因子和治疗靶点。本研究旨在探讨 circ_0088046 在 HCC 进展中的作用和机制。采用实时定量聚合酶链反应(qRT-PCR)、western blot 和免疫组织化学检测 circ_0088046、miR-1299、Rhotekin 2(RTKN2)、Bax、Bcl-2、E-钙黏蛋白和 Ki-67 的 mRNA 和蛋白表达。通过 5-乙炔基-2'-脱氧尿苷(EdU)检测和细胞集落形成检测来检测细胞增殖。通过流式细胞术测量细胞凋亡率。采用 Transwell 迁移和侵袭检测评估细胞迁移和侵袭。通过双荧光素酶报告基因检测和 RNA 免疫沉淀检测分析 miR-1299 与 circ_0088046 或 RTKN2 之间的分子靶关系。进行动物实验以证明 circ_0088046 对体内肿瘤形成的影响。HCC 组织和细胞中显示 circ_0088046 和 RTKN2 水平升高,miR-1299 水平降低。circ_0088046 缺失抑制 HCC 细胞增殖、迁移和侵袭,但促进细胞凋亡。miR-1299 是 circ_0088046 的靶标,miR-1299 抑制剂恢复了 circ_0088046 沉默对 HCC 细胞恶性的抑制作用。miR-1299 可以直接靶向 RTKN2,而过表达 RTKN2 挽救了 miR-1299 模拟物引起的抑制作用。此外,circ_0088046 沉默抑制体内肿瘤形成。circ_0088046 通过调节 miR-1299/RTKN2 轴促进 HCC 细胞恶性。

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