Li Haitao, Zhi Yunlai, Ma Chunyan, Shen Qianqian, Sun Fanghu, Cai Chengkuan
Department of Urology, Lianyungang Clinical College of Nanjing Medical University/The First People's Hospital of Lianyungang, Lianyungang, Jiangsu, People's Republic of China.
Cancer Manag Res. 2020 Nov 17;12:11701-11712. doi: 10.2147/CMAR.S273826. eCollection 2020.
Prostate cancer (PCa) is a major contributor to reduce the life quality of males. Circular RNAs were frequently reported to be associated with cancers. In the case of radiotherapy to PCa, the role of circ_0062020 was still inconclusive, which was further explored in this study.
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of circ_0062020, miR-615-5p and thyroid hormone receptor interactor 13 (TRIP13) in PCa tissues and cells, as well as in normal tissues and cell. Meanwhile, the proliferation of PCa cells was evaluated by clone formation assay and cell counting kit 8 (CCK8) assay. Moreover, the metastasis of PCa cells was assessed by transwell and wound healing assays. Furthermore, the apoptosis of PCa cells was determined by flow cytometry assay. Besides, dual-luciferase reporter system was applied to verify the correlation between miR-615-5p and circ_0062020 or TRIP13, which was predicted by online tool CircRNA interactome or TargetScan. In addition, the protein expression of TRIP13 was measured by Western blot in PCa tissues and cells and normal tissues and cells. Finally, xenograft tumor assay was performed to further confirming the function of circ_0062020 in PCa in vivo.
Circ_0062020 and TRIP13 were upregulated, while miR-615-5p was downregulated in PCa tissues and cells. Circ_0062020 knockdown or miR-615-5p overexpression inhibited the proliferation and metastasis, and promoted apoptosis, which could be reversed by miR-615-5p inhibitor or pc-TRIP13 in ionizing radiation (IR)-treated PCa cells. As expected, circ_0062020 sponged miR-615-5p to regulate TRIP13 expression in PCa cells. Circ_0062020 knockdown also suppressed PCa tumor growth in vivo.
Circ_0062020 suppressed the radiosensitivity by miR-615-5p/TRIP13 axis in PCa cells, which might provide insights into the radiotherapy for PCa.
前列腺癌(PCa)是降低男性生活质量的主要因素。环状RNA经常被报道与癌症有关。在PCa放疗的情况下,circ_0062020的作用仍不明确,本研究对此进行了进一步探讨。
采用定量实时聚合酶链反应(qRT-PCR)检测circ_0062020、miR-615-5p和甲状腺激素受体相互作用蛋白13(TRIP13)在PCa组织和细胞以及正常组织和细胞中的表达。同时,通过克隆形成试验和细胞计数试剂盒8(CCK8)试验评估PCa细胞的增殖。此外,通过Transwell和伤口愈合试验评估PCa细胞的转移。此外,通过流式细胞术检测PCa细胞的凋亡。此外,应用双荧光素酶报告系统验证miR-615-5p与circ_0062020或TRIP13之间的相关性,这是由在线工具CircRNA interactome或TargetScan预测的。此外,通过蛋白质免疫印迹法检测PCa组织和细胞以及正常组织和细胞中TRIP13的蛋白表达。最后,进行异种移植瘤试验以进一步证实circ_0062020在体内PCa中的功能。
在PCa组织和细胞中,circ_0062020和TRIP13上调,而miR-615-5p下调。circ_0062020敲低或miR-615-5p过表达抑制增殖和转移,并促进凋亡,在电离辐射(IR)处理的PCa细胞中,miR-615-5p抑制剂或pc-TRIP13可逆转这种作用。正如预期的那样,circ_0062020在PCa细胞中通过吸附miR-615-5p来调节TRIP13的表达。circ_0062020敲低也抑制了体内PCa肿瘤的生长。
circ_0062020通过miR-615-5p/TRIP13轴抑制PCa细胞的放射敏感性,这可能为PCa的放疗提供思路。