Circ_0037866通过分离miR-384提高Chromobox 5表达参与肾细胞癌的发生

Xiaoqiang Shi, Shichao Song, Ying Gao, Zhenyu Cui, Wentao Wang, Mingkai Liu
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引用次数: 1

摘要

背景:环状rna (circRNAs)已被证明在肾细胞癌(RCC)的癌变过程中发挥作用。因此,本研究旨在确定circ_0037866在调控RCC进展中的功能和分子机制。方法:采用实时定量聚合酶链反应和Western blotting检测基因和蛋白水平。体外实验包括集落形成、5-乙基-2 ' -脱氧尿苷、流式细胞术、transwell实验和体内肿瘤形成,研究circ_0037866对RCC肿瘤发生的影响。采用双荧光素酶报告基因法、RNA下拉法和RNA免疫沉淀法确认miR-384与circ_0037866或Chromobox 5 (CBX5)之间的相互作用。结果:Circ_0037866是一个稳定的circRNA,在RCC组织和细胞中被发现增加。在功能上,circ_0037866沉默抑制了RCC细胞的体外存活、侵袭和迁移,并在皮下异种移植模型中阻碍了RCC细胞的肿瘤发生。在机制上,circ_0037866可以作为miR-384的海绵,提高其靶CBX5的表达。此外,一系列挽救性实验表明,miR-384抑制逆转了circ_0037866敲低对RCC细胞的抗癌作用;此外,miR-384的恢复抑制了RCC细胞的生长和移动性,CBX5过表达减弱了这种抑制作用。结论:Circ_0037866敲低可通过miR-384/CBX5抑制RCC的肿瘤发生,为RCC治疗提供了一个有前景的分子靶点。
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Circ_0037866 Contributes to the Tumorigenesis of Renal Cell Carcinoma by Sequestering miR-384 to Elevate Chromobox 5 Expression
Background: Circular RNAs (circRNAs) were demonstrated to have roles in the carcinogenesis of renal cell carcinoma (RCC). Hence, this work aimed to determine the functions and molecular mechanism of circ_0037866 in regulating the progression of RCC. Methods: Quantitative real-time polymerase chain reaction and Western blotting were used to detect the levels of genes and proteins. In vitro assays, including colony formation, 5-ethynyl-2′-deoxyuridine, flow cytometry, transwell assays, and in vivo tumor formation, were conducted to investigate the effects of circ_0037866 on RCC tumorigenesis. Dual-luciferase reporter assay, RNA pull-down, and RNA immunoprecipitation assay were used to confirm the interaction between miR-384 and circ_0037866 or Chromobox 5 (CBX5). Results: Circ_0037866 is a stable circRNA and was found to be increased in RCC tissues and cells. Functionally, circ_0037866 silencing suppressed RCC cell survival, invasion, and migration in vitro, and impeded RCC cell tumorigenesis in the subcutaneous xenograft model. Mechanistically, circ_0037866 could function as a sponge for miR-384 to elevate the expression of its target CBX5. Furthermore, a series of rescue experiments showed that miR-384 inhibition reversed the anticancer effects of circ_0037866 knockdown on RCC cells; besides that, miR-384 restoration suppressed RCC cell growth and mobility, which were attenuated by CBX5 overexpression. Conclusion: Circ_0037866 knockdown restrains the tumorigenesis of RCC by miR-384/CBX5, revealing a promising molecular target for RCC therapy.
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