Anti-Tumor Effect of Celastrol on Hepatocellular Carcinoma by the circ_SLIT3/miR-223-3p/CXCR4 Axis

Abstract

Background 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. Methods 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. Results 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. Conclusion 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.

Circular RNA circ-BNC2 (hsa_circ_0008732) inhibits the progression of ovarian cancer through microRNA-223-3p/ FBXW7 axis

“…In this study, miR-223-3p was confirmed to be a target of circ-BNC2. It is reported that miR-223-3p plays different roles in different human malignancies [ 31 ]. For example, in non-small cell lung cancer, miR-223-3p restrains the malignant biological behaviors of tumor cells and induces the apoptosis via directly targeting ras homolog family member B [ 32 ].…”

Celastrol and Melatonin Modify SIRT1, SIRT6 and SIRT7 Gene Expression and Improve the Response of Human Granulosa-Lutein Cells to Oxidative Stress

“…The addition of celastrol, alone or with the addition of OS inductors increased SIRT7 gene expression under the treatment conditions tested in these experiments. Several studies point to celastrol as a substance with multiple activities such as anti-tumor [ 50 , 51 ], anti-inflammatory [ 52 , 53 ] and antioxidant [ 54 , 55 ], and it participates in cell processes as a molecule affecting signaling of several pathways such as the ERK pathway [ 56 , 57 ]. In this sense, celastrol has been reported to bind Shoc2, a scaffold protein involved in processes such as cell motility, invasion, and proliferation through the ERK pathway [ 58 ].…”

Celastrol: An Update on Its Hepatoprotective Properties and the Linked Molecular Mechanisms

“…As show in Table 5 ( Chen et al, 2011 ; Rajendran et al, 2012 ; Wei et al, 2014 ; Chang et al, 2016 ; Kun-Ming et al, 2020 ; Saber et al, 2020 ; Si et al, 2021 ), studies have manifested that celastrol exhibits anticancer activity against a variety of liver cancer animal models such as HCC patient-derived xenografts BALB/cJ mice ( Wei et al, 2014 ), different hepatocellular carcinoma cells derived xenografts mouse models ( Rajendran et al, 2012 ; Ma et al, 2014 ; Ren et al, 2017 ; Si et al, 2021 ), and DEN induced HCC rats/mice ( Chang et al, 2016 ; Saber et al, 2020 ) with dosage of 1–10 mg/kg for 3–10 weeks. Celastrol (4 mg/kg) prevented tumor proliferation and increased apoptosis via inhibition of STAT3/JAK2 signaling pathway in PLC/PRF5 cells derived xenografts HCC mice ( Rajendran et al, 2012 ).…”

“…Celastrol (4 mg/kg) prevented tumor proliferation and increased apoptosis via inhibition of STAT3/JAK2 signaling pathway in PLC/PRF5 cells derived xenografts HCC mice ( Rajendran et al, 2012 ). Recently, different studies validated that inhibition of circ_SLIT3/miR-223-3p/CXCR4 signaling is involved in the anti-HCC activity of celastrol both in vitro and in vivo ( Kun-Ming et al, 2020 ; Si et al, 2021 ). Meanwhile, celastrol has been demonstrated to decrease the hepatic lesions and elevation of serum alanine aminotransferase (ALT), glutamic oxalacetic transaminase (AST), alkaline phosphatase (ALP), and alpha fetoprotein (AFP) in diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) rats, due to activation of mitochondrial apoptosis induced by p53 ( Chang et al, 2016 ; Saber et al, 2020 ).…”