端锚聚合酶抑制剂XAV939抑制人骨肉瘤SOSP-9607细胞增殖及作用机制研究

Study on Inhibitory Effect and Its Mechanism of Tankyrase Inhibitor XAV939 on the Generation of Human Osteosarcoma SOSP-9607 Cells

目的:研究端锚聚合酶抑制剂XAV939对人骨肉瘤SOSP-9607细胞增殖的抑制及其作用机制。方法:以人骨肉瘤SOSP-9607细胞为研究对象,采用CCK-8法测定0.5、1、5、10μmol/L XAV939分别作用24、48、72 h后的细胞增殖情况,并计算细胞抑制率;采用流式细胞术检测5μmol/L XAV939作用72 h后的细胞凋亡情况及周期分布;以甘油醛-3-硫酸脱氢酶(GAPDH)为内参,采用蛋白质印迹法测定0.5、1、5、10μmol/L XAV939作用72 h后细胞中β-联蛋白(β-catenin)、G1/S期特异性周期蛋白D1(Cyclin D1)和基质金属蛋白酶7(MMP-7)的相对表达量。结果:1μmol/L XAV939作用72 h后,5、10μmol/L XAV939作用24、48、72 h后,SOSP-9607细胞的抑制率均显著升高(P<0.05或P<0.01)。5μmol/L XAV939作用72 h后,SOSP-9607细胞的凋亡率从3.71%上升至21.03%(P<0.05);G1期细胞的比例由45.5%增至54.8%,S期细胞的比例由27.4%降至24.0%,G2/M期细胞的比例由22.2%降至16.2%(P<0.05)。5、10μmol/L XAV939作用72 h后,SOSP-9607细胞中β-catenin、Cyclin D1和MMP-7的相对表达量均显著下降(P<0.05或P<0.01)。结论:端锚聚合酶抑制剂XAV939能够抑制人骨肉瘤SOSP-9607细胞的增殖,其机制可能与其将细胞周期阻滞于G1期、阻断细胞外因子(Wnt)/β-catenin信号通路有关。

OBJECTIVE:To study the inhibitory effect of tankyrase inhibitor XAV939 on human osteosarcoma SOSP-9607 cells and its mechanism.METHODS:The human osteosarcoma SOSP-9607 cells were selected as research objects.The proliferation of osteosarcoma cells was determined by CCK-8 method after treated with 0.5,1,5,10μmol/L XAV939 for 24,48,72 h,and the inhibitory rate was calculated.After treated with 5μmol/L XAV939 for 72 h,flow cytometry was applied to detect cell apoptosis and cell cycle distribution of osteosarcoma cells.Using glyceraldehyde-3-dehydrogenase(GAPDH)as internal reference,Western blot assay was used to detect the relative expression ofβ-catenin,Cyclin D1 and MMP-7 in osteosarcoma cells after treated with 0.5,1,5,10μmol/L XAV939 for 72 h.RESULTS:After treated with 1μmol/L XAV939 for 72 h,5,10μmol/L XAV939 for 24,48,72 h,the inhibitory rates of SOSP-9607 cells were increased significantly(P<0.05 or P<0.01).After treated with 5μmol/L XAV939 for 72 h,the apoptotic rate of SOSP-9607 cells was increased from 3.71%to 21.03%(P<0.05);the proportion of G1-phase cells increased from 45.5%to 54.8%,that of S-phase cells decreased from 27.4%to 24.0%,that of G2/M-phase cells decreased from 22.2%to 16.2%(P<0.05).After treated with 5,10μmol/L XAV939 for 72 h,the relative expression ofβ-catenin,Cyclin D1 and MMP-7 in SOSP-9607 cells decreased significantly(P<0.05 or P<0.01).CONCLUSIONS:Tankyrase inhibitor XAV939 can inhibit the proliferation of human osteosarcoma SOSP-9607 cells,the mechanism of which may be associated with arresting cells at G1 phase and blocking signaling pathway of Wnt/β-catenin.