Effect of catalpol on the malignant biological behavior of oral squamous cell carcinoma cells by regulating the Keap1/Nrf2/HO-1 signaling pathway
Objective To explore the effect of catalpol on the malignant biological behaviors of oral squamous cell carcinoma cells by regulating the kelch-like ECH-associated protein 1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods Oral squamous cell carcinoma cells Tac8113 were cultured in vitro.The optimal treatment concentration of catalpol was determined by CCK-8 assay.Tac8113 cells were induced with blank control,24μg/mL catalpol,transfection of negative control shRNA(sh-NC),transfection of Keap1 shRNA(sh-Keap1),24μg/mL catalpol+transfection of sh-NC and 24μg/mL catalpol+transfection of sh-Keap1.Cell proliferation,migration and apoptosis were detected by CCK-8 assay,wound healing assay and flow cytometry were performed,respectively.Reactive oxygen species(ROS)level was measured using the dichloro-dihydro-fluorescein diacetate(DCFH-DA)staining.Superoxide dismutase(SOD)and malondialdehyde(MDA)levels were measured using commercial kits.Protein expressions of proteins in the Keap1/Nrf2/HO-1 signaling pathway and apoptotic proteins were examined by Western blot.Results Compared with that of blank control,the survival rate of Tac8113 cells dose-dependently decreased with the treatment of catalpol(P<0.05),and at last,24μg/mL catalpol was adopted in the following analyses.Compared with those induced with blank control,Tac8113 cells induced with 24μg/mL catalpol presented significantly lower OD450,wound healing rate,ROS and MDA levels and protein expressions of Bcl-2,Nrf2 and HO-1,but significantly higher apoptotic rate,SOD level and protein expressions of Keap1 and caspase 3(P<0.05).Knockdown of Keap1 aggravated the malignant behaviors of Tac8113 cells,and reversed the regulatory effects of catalpol on the malignant behaviors of Tac8113 cells(P<0.05).Conclusion Catalpol exerts the anti-tumor effect on oral squamous cell carcinoma cells by inhibiting the proliferation and migration and inducing cell apoptosis via upregulating Keap1 and downregulating Nrf2 and HO-1.
万方数据
维普
