淫羊霍苷对人骨肉瘤细胞U2OS增殖、凋亡和迁移的调节作用

Effects of Icariin on Cell Proliferation, Apoptosis and Migration of Human Osteosarcoma Cell Line U2OS

目的探究淫羊霍苷(Icariin)对人骨肉瘤细胞U20S增殖、凋亡和迁移的作用。方法将人骨肉瘤细胞系U20S细胞随机分为U2OS组,Icariin(5μmol/L)组、Icariin(10μmol/L)组和Icariin(20μmol/L)组。用5、10和20μmol/L的淫羊霍苷分别处理U2OS细胞0、1、2、3和4d,CCK8法检测细胞活性;用不同浓度淫羊霍苷处理U2OS细胞24h,流式细胞术检测U2OS细胞凋亡情况,划痕实验检测各组细胞的迁移能力,免疫印迹检测Ki67、eleaved Caspase-3和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达。结果根据预实验结果选择5、10和20μmol/L3个浓度进行后续实验。与U2OS组比较,Icariin(5μmol/L)组、Icariin(10μmol/L)组和Icariin(20μmol/L)组细胞生长速度明显降低,增殖相关蛋白Ki67的表达水平也明显降低;同时,Icariin(5(μmol/L)组、Icariin(10μmol/L)组和Icariin(20μmol/L)组细胞凋亡率与U2OS组比较明显升高,凋亡标记蛋白cleaved Caspase-3的表达水平也显著升高;此外,与U2OS组比较,Icariin(10μmol/L)组和Icariin(20μmol/L)组细胞划痕闭合率明显降低,迁移相关蛋白MMP-9的表达也明显被抑制。结论淫羊霍苷能抑制人骨肉瘤细胞系U2OS细胞增殖和迁移,并诱导U2OS细胞凋亡。

Objective To inveastigate the effect oilcariinon cell proliferation, apoptosis and migration of human osteosarcoma cell line U2OS. Methods Cells were divided into U2OS group,Icariin (5 μmol/L) group, Icariin ( 10 μmol/L) group and Icariin (20 μmoll/L) group. Cells were treated with icariin (at 0, 5 , 10, 20 μmol/L) for 0, 1,2,3 and 4d respectively, and cell viability was measured by CCK8 assay. Cells were treated with different doses of Icariin for 24 h. Flow cytometry was perfbnned to determine cell apoptosis;the protein levels of Ki67 , cleaved caspase-3 and matrix metalloproteinase-9 ( MMP-9) were measured by Western blotting. Results On the basis of preliminary study, 5 μmol/L, 10 μmol/L and 20μmol/L were selected for further study. Compared with U2OS group, the growth of U2OS cells in Icariin (5 μmol/L) group, Icariin ( 10 μmol/L) group and Icariin (20 μmol/L) group were decreased significantly , and the expression level of Ki67 was down-regulated as compared with U2OS group. Meanwhile, the cell apoptosis nites of cells in Icariin (5 μmol/L), Icariin ( 10 μmol/L) and Icariin (20 μmol/L) groups were increased when compared with U2OS group, and the protein level of apoptosis-rated protein ,cleaved caspase-3 in Icariin ( 5 μmol/L), Icariin ( 10 μmol/L) and Icariin ( 20 μmol/L) groups was increased remarkedly compared with U2OS group. In addition, compared with U2OS group, the wound closure rate of cells and the expressions of migration-related protein , MM P-9 in Icariin (5 μmol/L), Icariin ( 10 μmol/L) and Icariin (20 μmol/L) groups were decreased significantly.Conclusion Icariin can inhibit the proliferation and migration, and induce cell apoptosis of human osteosarcoma cell line U2OS.