【摘要】 目的 探讨川楝素(TSN)调控微小RNA-409-3p(miR-409-3p)对前列腺癌细胞增殖、迁移、侵袭及血管内皮生长因子(VEGF)/血管内皮生长因子受体2(VEGFR2)通路的影响。方法 使用0、10、20、40、80和160μmol/L的TSN处理PC3细胞，采用噻唑蓝法检测PC3细胞存活率，选择最佳药物浓度。将细胞分为对照组(Control组)、TSN低浓度组(TSN-L组)、TSN中浓度组(TSN-M组)、TSN高浓度组(TSN-H组)、TSN高浓度+miR-409-3p拮抗剂阴性对照组(TSN-H+antagomiR-NC组)、TSN高浓度+miR-409-3p拮抗剂组(TSN-H+antagomiR-409-3p组)。采用实时荧光定量聚合酶链反应(qRT-PCR)检测PC3细胞中miR-409-3p表达；EdU法检测PC3细胞增殖水平；Transwell小室实验和划痕愈合实验检测PC3细胞的侵袭和迁移能力；Western blot检测细胞周期蛋白D1(CyclinDl)、细胞周期蛋白依赖性激酶4(CDK4)、VEGF、VEGFR2蛋白表达。结果 与0μmol/L比较，10、20、40、80和160μmol/L的TSN细胞存活率显著降低(P<0.05),选择20、40、80μmol/L的TSN用于后续实验。与Control组比较，TSN-L组、TSN-M组和TSN-H组PC3细胞EdU阳性率、细胞侵袭数、划痕愈合率、CyclinDl、CDK4、VEGF、VEGFR2蛋白表达显著降低(P<0.05),miR-409-3p表达显著增加(P<0.05),且呈浓度依赖性。与TSN-H+antagomiR-NC组比较，TSN-H+antagomiR-409-3p组PC3细胞EdU阳性率、细胞侵袭数、划痕愈合率、CyclinDl、CDK4、VEGF、VEGFR2蛋白表达显著增加(P<0.05),miR-409-3p表达显著降低(P<0.05)。结论 TSN通过上调miR-409-3p表达抑制VEGF/VEGFR2通路的激活，进而抑制PC3细胞增殖、迁移、侵袭。更多还原

【Abstract】 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.更多还原