The Inhibitory Effect And Molecular Mechanism Of Jatrorrhizine Hydrochloride On Prostate Cancer

Objective: To investigate the effects of jatrorrhizine hydrochloride on proliferation,plate clone formation,migration,invasion and apoptosis of prostate cancer cells.To explore the effect of jatrorrhizine hydrochloride on subcutaneous tumor-bearing nude mice at the animal level,and to further explore its molecular mechanism.Methods: Prostate cancer PC-3 cells were cultured in vitro,and the binding effect between drug and protein complex was investigated by molecular docking.The effects of different concentrations of jatrorrhizine hydrochloride on the proliferation of PC-3 cells at 24 h and48 h were detected by MTT assay,and the effect of jatrorrhizine hydrochloride on the clone formation of prostate cancer PC-3 cells was detected by plate clone formation assay.Cell wound healing test and Transwell cell migration assay were used to detect the effect of jatrorrhizine hydrochloride on the transverse and longitudinal migration of PC-3 cells.Transwell cell invasion assay was used to detect the effect of jatrorrhizine hydrochloride on the invasive ability of PC-3 cells.Hochest 33342 dye was used to detect whether jatrorrhizine hydrochloride promoted the apoptosis of PC-3 cells.PC-3 cells were inoculated subcutaneously into nude mice to establish the subcutaneous tumor-bearing model of nude mice.The effect of jatrorrhizine hydrochloride on the proliferation of tumor-bearing nude mice was observed by comparing with the blank control group.HE staining and immunohistochemical experiments were used to verify whether jatrorrhizine hydrochloride has toxic reaction on organs in vivo and its possible mechanism.The effects of different concentrations of jatrorrhizine on the expression of EMT related proteins in PC-3 cells and the expression of oncogene MTA1 were detected by Western blot.Results: MTT and plate cloning assay showed that jatrorrhizine hydrochloride could inhibit prostate cancer PC-3 cells in a time and dose dependent manner,and wound healing and Transwell migration and invasion tests showed that jatrorrhizine hydrochloride could inhibit the horizontal and vertical migration and invasion of PC-3 cells in a dose-dependent manner.Hochest 33342 apoptosis staining showed that jatrorrhizine hydrochloride could promote apoptosis of PC-3 cells in a concentration-dependent manner.The results of subcutaneous tumor loading in nude mice showed that the growth rate of subcutaneous tumor in the jatrorrhizine hydrochloride group was significantly slower than that in the blank control group,and the tumor volume was smaller than that in the blank control group,indicating that jatrorrhizine hydrochloride inhibited the tumor growth rate of PC-3 cells in nude mice.The results of HE staining and immunohistochemistry showed that jatrorrhizine hydrochloride did not show obvious toxicity to the heart,liver,spleen,lung and kidney of nude mice and had a certain effect on the expression of EMT marker protein.In addition,the molecular docking results showed that there was a strong binding between jatrorrhizine hydrochloride and MTA1 protein complex.The results of Western blot showed that compared with the blank control group,the expression of MTA1 in the treatment group was significantly downregulated,and the downward trend was more obvious with the increase of drug concentration.In addition,jatrorrhizine hydrochloride could inhibit the expression of N-cadherin snail,a key protein in EMT,and upregulate the expression of E-cadherin.Conclusion: This experiment shows that through cell experiments in vitro and animal experiments in vivo,jatrorrhizine hydrochloride can inhibit the EMT progression of prostate cancer PC-3 cells by binding to MTA1 protein complex and inhibit the proliferation,invasion and migration of prostate cancer cells without obvious side effects.