The effect of toosendanin on the proliferation,migration,invasion and vascular endothelial growth factor/vascular endothelial growth factor receptor 2 pathway of prostate cancer cells by regulating microRNA-409-3p
Objective To investigate the effects of toosendanin(TSN)on the proliferation,migration,invasion and vascular endothelial growth factor/vascular endothelial growth factor receptor 2(VEGF/VEGFR2)pathway of prostate cancer cells by regulating microRNA-409-3p(miR-409-3p).Methods PC3 cells were treated with TSN of 0,10,20,40,80,and 160 μmol/L,and the survival rate of PC3 cells was detected by thiazole blue method,and the optimal drug concentration was selected.The cells were divided into control group,low concentrations toosendanin group(TSN-L group),medium concentrations toosendanin group(TSN-M group),high concentrations toosendanin group(TSN-H group),high concentration TSN+miR-409-3p antagonist negative control group(TSN-H+antagomiR-NC group),high concentration TSN+miR-409-3p antagonist group(TSN-H+antagomiR-409-3p group).Real-time quantitative fluorescent polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-409-3p in PC3 cells;EdU method was used to detect the proliferation level of PC3 cells;Transwell chamber experiment and scratch healing experiment were used to detect the abilities of invasion and migration in PC3 cells;Western blot was used to detect the expression of CyclinDl,cyclin dependent kinase 4(CDK4),VEGF,and VEGFR2 proteins.Results Compared with 0 µmol/L,the survival rate of PC3 cells treated with 10,20,40,80 and 160 µmol/L TSN was significantly reduced(P＜0.05),20,40,80 µmol/L TSN were selected for subsequent experiments.Compared with the control group,the EdU positive rate,number of cell invasion,scratch healing rate,the expressions of CyclinDl,CDK4,VEGF,and VEGFR2 protein of PC3 cells in the TSN-L group,TSN-M group,and TSN-H group were significantly reduced(P＜0.05),the expressions of miR-409-3p were significantly increased(P＜0.05),with a concentration dependent manner.Compared with the TSN-H+antagomiR-NC group,the EdU positive rate,number of cell invasion,scratch healing rate,the expressions of CyclinDl,CDK4,VEGF,and VEGFR2 protein of PC3 cells in the TSN-H+antagomiR-409-3p group were significantly increased(P＜0.05),the expressions of miR-409-3p were significantly reduced(P＜0.05).Conclusions TSN inhibits the activation of the VEGF/VEGFR2 pathway by up-regulating the expression of miR-409-3p,thereby inhibiting the proliferation,migration,and invasion of PC3.
NETL
NSTL
万方数据
