染料木黄酮对骨肉瘤细胞增殖及侵袭的影响及作用机制

目的 探讨染料木黄酮对人骨肉瘤MG-63细胞增殖、侵袭的影响及其作用机制.方法 用染料木黄酮处理MG-63细胞,CCK-8法检测细胞增殖；Transwell小室体外侵袭实验法检测MG-63细胞侵袭能力；实时定量聚合酶链反应(PCR)和蛋白印迹法(Western blot)分别检测MG-63细胞基质金属蛋白酶(MMP)-2／MMP-9的mRNA及蛋白水平.结果CCK-8法表明染料木黄酮处理组MG-63细胞增殖被明显抑制,抑制率最高(85.3±5.1)％(P＜0.05)；染料木黄酮在0、12.5、25.0、50.0 μmol/L时穿膜细胞数分别为(96.5±5.5)、(88.4±3.2)、(53.8±4.4)、(36.6±3.8)个,可明显降低MG-63细胞的侵袭能力(P＜0.05)；染料木黄酮处理后MMP-2及MMP-9的mRNA表达水平明显下降(P＜0.05)；MMP-2/MMP-9蛋白表达水平也明显下降,且具有时间依赖性.结论染料木黄酮抑制骨肉瘤MG-63细胞增殖及侵袭转移能力,这种作用与MMP-2及MMP-9表达变化有关.

Effects of Genistein on proliferation and invasion of osteosarcoma cells and action mechanism

Objective To evaluate the effect of Genistein on the proliferation and invasion of osteosarcoma cells.Methods CCK-8 was used to detect the proliferation of osteosarcoma cells treated with different concentrations of Genistein.Transwell invasion assay was used to measure the invasion of osteosarcoma cells.The matrix metalloproteinase (MMP)-2/9 expression was analyzed by Western blotting and real-time quantitative-polymerase chain reaction (PCR) in the MG-63 osteosarcoma cells treated with Genistein.Results CCK-8 showed that Genistein could inhibit the proliferation of osteosarcoma cells in a time- and dose-independent manner with the highest inhibition ratio being ( 85.3 ± 5.1 ) ％ ( P ＜ 0.05 ).Transwell invasion assays revealed that the number of cells was (96.5 ± 5.5 ),( 88.4 ± 3.2),( 53.8 ±4.4),(36.6 ±3.8) at the concentrations of 0,12.5,25.0,50.0 μmol/L Genistein respectively,suggesting Genistein could significantly inhibit the invasion of osteosarcoma cells ( P ＜ 0.05 ).MMP-2/9 mRNA and protein expression was significantly inhibited by Genistein ( P ＜ 0.05 ) in a time-independent manner.Conclusion Genistein can inhibit proliferation and invasion of osteosarcoma cells in vitro,which is associated with the changes of MMP-2/9 expression.