JG-231对结直肠癌细胞恶性表型的影响及其作用机制OA

OBJECTIVE:To investigate effects of the small molecule inhibitor JG-231 on malignant phenotype of colorectal cancer(CRC)cells and to explore its mechanisms of action.METHODS:Colorectal cancer cell lines HCT116 and DLD-1 were treated with JG-231 at various concentrations(0.5-8 μmol/L)for 24 or 48 hours,with DMSO serving as the control.Cell viability was measured using the CCK-8 assay;cell proliferation was evaluated by the plate colony formation assay;cell cycle distribution and apoptosis were analyzed by flow cytometry;expression levels of the AKT signaling pathway and cell cycle/apoptosis-related proteins were detected by Western blot.Furthermore,a subcutaneous xenograft model was established by inoculating DLD-1 cells into the right flank of nude mice to evaluate inhibitory effect of JG-231 on tumor growth in vivo.RESULTS:Compared with the control group,JG-231 treatment significantly inhibited viability of HCT116 and DLD-1 cells(P<0.01).The colony formation ability of the JG-231-treated groups was significantly lower than that of the control group(P<0.01).Flow cytometry results demonstrated that JG-231 significantly induced G0/G1 phase arrest and apoptosis in both HCT116 and DLD-1 cells(P<0.01).Molecular analysis revealed that the total and phosphorylated protein levels of AKT and its downstream molecule GSK-3β were significantly downregulated following JG-231 treatment.Meanwhile,expression of the anti-apoptotic protein Mcl-1 and the cell cycle protein Cyclin D1 decreased,whereas the levels of apoptotic markers cleaved Caspase-3 and cleaved PARP1 increased.In vivo experiments showed that JG-231 significantly inhibited the volume and weight of xenograft tumors in nude mice compared to the control group(P<0.01),with no observed systemic toxicity.CONCLUSION:JG-231 significantly inhibited proliferation and colony formation of colorectal cancer cells both in vitro and in vivo,and induced G0/G1 phase arrest and apoptosis.These effects may be attributed to inhibition of the AKT signaling pathway and subsequent regulation of downstream cell cycle and apoptosis-related proteins.