Si Hailong, Wang Huiling, Xiao Haijuan, Fang Yu, Wu Zhaoli
First School of Clinical Medical, Shaanxi University of Traditional Chinese Medicine, Xianyang, 712000, People's Republic of China.
Department of Oncology, Affiliated Hospital of the Shaanxi University of Traditional Chinese Medicine, Xianyang, 712000, People's Republic of China.
Cancer Manag Res. 2021 Feb 5;13:1099-1111. doi: 10.2147/CMAR.S278023. eCollection 2021.
Celastrol is a potential anti-tumor agent in hepatocellular carcinoma (HCC). Identifying the molecular determinants of the anti-HCC effect of celastrol is still challenging. In this study, we undertook to associate circular RNAs (circRNAs) with the anti-HCC molecular determinants of celastrol.
Cell colony formation, proliferation, migration, invasion and apoptosis were determined using the colony formation, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTS), transwell and flow cytometry assays, respectively. The levels of circRNA slit guidance ligand 3 (circ_SLIT3), miR-223-3p and C-X-C motif chemokine receptor 4 (CXCR4) were gauged by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Ribonuclease R (RNase R) and actinomycin D assays were performed to assess the stability of circ_SLIT3. Targeted relationships among circ_SLIT3, miR-223-3p and CXCR4 were confirmed by the dual-luciferase reporter assay. In vivo assays were performed to detect the roles of celastrol and circ_SLIT3 on tumor growth in vivo.
Celastrol repressed HCC cell proliferation, migration, invasion, and enhanced apoptosis in vitro and suppressed tumor growth in vivo. Celastrol down-regulated circ_SLIT3 expression in HCC cells, and celastrol exerted an anti-tumor effect on HCC in vitro and in vivo by down-regulating circ_SLIT3. Mechanistically, circ_SLIT3 directly interacted with miR-223-3p, and circ_SLIT3 controlled CXCR4 expression by sponging miR-223-3p. Moreover, miR-223-3p was involved in the celastrol/circ_SLIT3-mediated regulation on HCC progression. Furthermore, celastrol exerted the anti-HCC effect in vitro through the miR-223-3p/CXCR4 axis.
Our present work first identified the circ_SLIT3/miR-223-3p/CXCR4 axis as a novel mechanism of the anti-HCC effect of celastrol, providing a new insight into the involvement of circRNAs in the anti-tumor molecular determinants of celastrol.
雷公藤红素是一种潜在的肝细胞癌（HCC）抗肿瘤药物。确定雷公藤红素抗HCC作用的分子决定因素仍然具有挑战性。在本研究中，我们致力于将环状RNA（circRNAs）与雷公藤红素的抗HCC分子决定因素联系起来。
分别使用集落形成、3-（4,5-二甲基噻唑-2-基）-2,5-二苯基-2H-四唑溴盐（MTS）、Transwell和流式细胞术检测细胞集落形成、增殖、迁移、侵袭和凋亡。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测环状RNA缝隙引导配体3（circ_SLIT3）、miR-223-3p和C-X-C基序趋化因子受体4（CXCR4）的水平。进行核糖核酸酶R（RNase R）和放线菌素D检测以评估circ_SLIT3的稳定性。通过双荧光素酶报告基因检测证实circ_SLIT3、miR-223-3p和CXCR4之间的靶向关系。进行体内实验以检测雷公藤红素和circ_SLIT3对体内肿瘤生长的作用。
雷公藤红素在体外抑制HCC细胞增殖、迁移、侵袭并增强凋亡，在体内抑制肿瘤生长。雷公藤红素下调HCC细胞中circ_SLIT3的表达，并且雷公藤红素通过下调circ_SLIT3在体外和体内对HCC发挥抗肿瘤作用。机制上，circ_SLIT3直接与miR-223-3p相互作用，并且circ_SLIT3通过海绵吸附miR-223-3p来控制CXCR4的表达。此外，miR-223-3p参与了雷公藤红素/circ_SLIT3介导的对HCC进展的调控。此外，雷公藤红素在体外通过miR-223-3p/CXCR4轴发挥抗HCC作用。
我们目前的工作首次确定circ_SLIT3/miR-223-3p/CXCR4轴是雷公藤红素抗HCC作用的新机制，为circRNAs参与雷公藤红素的抗肿瘤分子决定因素提供了新的见解。
