【摘要】 目的探讨AG1478对卵巢癌细胞生物活性及纤溶酶原激活物抑制物1(PAI-1)的作用机制。方法将CaOV-3细胞分为空白组和干预组,空白组为无药物干预的CaOV-3细胞,干预组分别为干预A组(5μmol的AG1478溶液)、干预B组(10μmol的AG1478溶液)、干预C组(15μmol的AG1478溶液)和干预D组(20μmol的AG1478溶液)。分别采用四甲基偶氮唑蓝(MTT)法、流式细胞仪及Transwell小室检测细胞抑制率、凋亡率及侵袭数目,采用蛋白质印迹(Western blot)法和逆转录聚合酶链反应(RT-PCR)分别检测PAI-1、尿激酶型纤溶酶原激活剂受体(uPAR)蛋白和mRNA水平。结果不同时间点干预B组、干预C组和干预D组CaOV-3细胞抑制率均高于干预A组,干预D组CaOV-3细胞抑制率均高于干预B组,差异均有统计学意义(P﹤0.05)。干预组CaOV-3细胞凋亡率均高于空白组,细胞侵袭数目均少于空白组,干预B组、干预C组、干预D组CaOV-3细胞凋亡率均高于干预A组,细胞侵袭数目均少于干预A组,干预D组CaOV-3细胞凋亡率高于干预B组,细胞侵袭数目少于干预B组,差异均有统计学意义(P﹤0.05)。干预组CaOV-3细胞中PAI-1、uPAR蛋白和mRNA表达量均低于空白组,干预B组、干预C组及干预D组CaOV-3细胞中PAI-1、uPAR蛋白和mRNA表达量均低于干预A组,干预D组CaOV-3细胞中PAI-1、uPAR蛋白和mRNA表达量均低于干预B组,差异均有统计学意义(P﹤0.05)。结论AG1478能够有效降低卵巢癌CaOV-3细胞存活率及侵袭性,加快凋亡,作用机制可能与阻碍PAI-1和uPAR信号通路表达有关。更多还原

【Abstract】 Objective To investigate the mechanism of AG1478 on the biological activity of ovarian cancer cells and plasminogen activator inhibitor type 1(PAI-1). Method The CaOV-3 cells were divided into blank group and intervention group. The blank group was CaOV-3 cells without drug intervention, and the intervention group was divided into intervention group A(5 μmol AG1478 solution), intervention group B(10 μmol AG1478 solution), intervention C group(15 μmol AG1478 solution), and intervention D group(20 μmol AG1478 solution). The cell inhibition rate, apoptosis rate, and the number of invasions were detected by the methyl thiazolyl terazolium(MTT) method, flow cytometer, and Transwell chamber assay. Western blot and reverse transcription-polymerase chain reaction(RT-PCR) were used to detect PAI-1, urokinase type plasminogen activator receptor(uPAR) protein and mRNA levels, respectively. Result The inhibitory rate of CaOV-3 cells in intervention group B, intervention group C, and intervention group D were higher than those in intervention group A at different time points, and the inhibitory rate of CaOV-3 cells in intervention group D was higher than that in intervention group B, and the differences were statistically significant(P<0.05). The apoptosis rate of CaOV-3 cells in the intervention groups were higher than those of the blank group, and the number of invasion cells were less than those of the blank group. The apoptosis rate of CaOV-3 cells in intervention group B, intervention group C, and the intervention group D were all higher than that of intervention group A, while the number of invasion cells were less than that of intervention group A, the rate of apoptosis of CaOV-3 cells in intervention group D was higher than that of intervention group B, the number of invasion cells was less than that of intervention group B, the differences were statistically significant(P<0.05). The expression levels of PAI-1, uPAR protein and mRNA in CaOV-3 cells in the intervention group were lower than those in the blank group, the expressions of PAI-1, uPAR protein and mRNA in CaOV-3 cells of intervention group B, intervention C, and intervention group D were lower than those of intervention group A, the expression of PAI-1, uPAR protein and mRNA in CaOV-3 cells of intervention group D was lower than the intervention group B, and the differences were statistically significant(P<0.05). Conclusion AG1478 could effectively reduce the survival rate of ovarian cancer CaOV-3 cells and inhibit their invasiveness while inducing apoptosis, the underlying mechanism may be related to blocking the expression of PAI-1 and relevant uPAR signaling pathways.更多还原