| Objective Pectolinarigenin(PEC),an active compound isolated from traditional herbal medicine,has shown potential anti-tumor properties against various types of cancer cells.However,its mechanism of action in bladder cancer,which is one of the fatal human carcinomas,remains unexplored.This research will provide insight into whether PEC can be used as a potential chemotherapeutic agent for bladder cancer and its specific mechanisms.Methods MTT and clone formation assays were used to evaluate cell proliferation rates.Cell cycle analysis was performed using flow cytometry.Protein levels were estimated using western blots,immunofluorescence,and immunohistochemistry.Gene expressions were measured using q RT-PCR.Comet assay was used to measure DNA damage.Transmission electron microscopy was used to observe cellular ultrastructure.Molecular docking and cellular thermal shift assay were performed to verify the target of PEC.BALB/c-nude mice were used to establish xenograft models.Results PEC effectively inhibited bladder cancer cell proliferation and caused cell cycle arrest in the G2/M phase.Further,molecular docking and cellular thermal shift assay revealed that PEC caused intracellular DNA damage by targeting TOP2A(DNA topoisomerase Ⅱ alpha).In addition,this study confirmed that DNA damage caused by PEC was induced by the p53 pathway in G2/M phase cell cycle arrest.Also,by exploring the changes of autophagic flow,we found that PEC could block the late process of intracellular autophagic flow,causing further DNA damage and cell proliferation inhibition.Finally,in vivo animal experiments demonstrated the effective carcinogenic effect and safety of PEC in vivo.Conclusions In this study,it was demonstrated that PEC can effectively inhibit bladder cancer cell proliferation by targeting TOP2 A to cause DNA damage and inhibiting autophagy,which is promising as a new chemotherapeutic agent for the treatment of bladder cancer. |
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