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Circ_0000370通过调控miR-502-5p/SIRT1轴在结直肠癌中发挥致癌作用。

Circ_0000370 Plays an Oncogenic Role in Colorectal Cancer by Regulating the miR-502-5p/SIRT1 Axis.

作者信息

Li Zhu, Tong Gangling, Peng Xiaodan, Wang Shubin

机构信息

Department of Oncology, Peking University Shenzhen Hospital, 1120 Lianhua Road, Futian District, Shenzhen, 518000, Guangdong, China.

出版信息

Biochem Genet. 2024 Apr;62(2):1231-1247. doi: 10.1007/s10528-023-10468-9. Epub 2023 Aug 10.

Abstract

The importance of circular RNA has been reported in cancer development. However, the role and mechanism of circ_0000370 in CRC progression are still unclear. Quantitative real-time PCR and Western blot assay were performed to measure RNA and protein expression. Cell proliferation was assessed by cell colony formation assay and 5-Ethynyl-2'-deoxyuridine assay. Flow cytometry was used to measure cell apoptosis. Cell migration and invasion were detected by transwell assay. The intermolecular target relations between miR-502-5p and circ_0000370 or SIRT1 were confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. A xenograft tumor model was established to examine the role of circ_0000370 in tumor growth in vivo. As compared with controls, the expression of circ_0000370 was upregulated in CRC tissues and cells. Circ_0000370 depletion inhibited CRC cell proliferation, migration and invasion but induced cell apoptosis. Meanwhile, circ_0000370 depletion restrained tumor growth in vivo. In addition, miR-502-5p inhibitor partly reverted the impacts of circ_0000370 knockdown on CRC cells. Moreover, miR-502-5p mimic-caused effects on cell phenotypes were attenuated by SIRT1 overexpression. Circ_0000370 induced the proliferation and metastasis of CRC cells by sponging miR-502-5p and enhancing SIRT1 expression, which provided a possible target for CRC treatment.

摘要

环状RNA在癌症发展中的重要性已有报道。然而,circ_0000370在结直肠癌进展中的作用和机制仍不清楚。采用定量实时聚合酶链反应和蛋白质免疫印迹法检测RNA和蛋白质表达。通过细胞集落形成试验和5-乙炔基-2'-脱氧尿苷试验评估细胞增殖。采用流式细胞术检测细胞凋亡。通过Transwell试验检测细胞迁移和侵袭。通过双荧光素酶报告基因试验和RNA免疫沉淀试验证实了miR-502-5p与circ_0000370或SIRT1之间的分子间靶向关系。建立异种移植肿瘤模型以研究circ_0000370在体内肿瘤生长中的作用。与对照组相比,circ_0000370在结直肠癌组织和细胞中的表达上调。circ_0000370的缺失抑制了结直肠癌细胞的增殖、迁移和侵袭,但诱导了细胞凋亡。同时,circ_0000370的缺失抑制了体内肿瘤的生长。此外,miR-502-5p抑制剂部分逆转了circ_0000370敲低对结直肠癌细胞的影响。此外,SIRT1的过表达减弱了miR-502-5p模拟物对细胞表型的影响。circ_0000370通过海绵吸附miR-502-5p并增强SIRT1表达诱导结直肠癌细胞的增殖和转移,这为结直肠癌的治疗提供了一个可能的靶点。

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