Li Zhu, Tong Gangling, Peng Xiaodan, Wang Shubin
Department of Oncology, Peking University Shenzhen Hospital, 1120 Lianhua Road, Futian District, Shenzhen, 518000, Guangdong, China.
Biochem Genet. 2024 Apr;62(2):1231-1247. doi: 10.1007/s10528-023-10468-9. Epub 2023 Aug 10.
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.
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。
