淫羊藿素对骨肉瘤细胞增殖分化及侵袭力的影响

Effect of Icaritin on the proliferation and differentiation and invasion of Osteosarcoma cells

目的研究淫羊藿素对骨肉瘤细胞增殖分化及侵袭力作用机制。方法以骨肉瘤细胞MG-63为研究对象,分别用4、8、16、32、64μmol/L浓度的淫羊藿素与骨肉瘤细胞MG-63细胞作用24 h、48 h、72 h,同时设置空白对照组,经MTT法检测细胞抑制情况。通过Annexin V/PI双染色淫羊藿素浓度为4、8、16、32μmol/L作用48 h后的骨肉瘤细胞MG-63经流式细胞仪观察并计算细胞凋亡率。Transwell小室检测经淫羊藿素浓度为4、8、16、32μmol/L作用48 h后的骨肉瘤细胞MG-63侵袭能力。Western blot检测淫羊藿素对细胞中Bax、Bcl-2、MMP-9表达影响。结果与对照组相比,淫羊藿素作用后的骨肉瘤细胞MG-63,抑制率有显著差异(P<0.05)。骨肉瘤细胞经0、4、8、16、32μmol/L淫羊藿素作用后的细胞凋亡率依次为0.125 1±0.018 4,0.241 4±0.019 7,0.436 5±0.030 1,0.723 2±0.031 5,0.789 7±0.047,进一步说明了淫羊藿素是通过促进癌细胞凋亡抑制癌细胞增殖的。浓度为4、8、16、32μmol/L淫羊藿素作用后的骨肉瘤MG-63细胞侵袭力与对照组相比差异显著(P<0.01)。淫羊藿素可以减弱骨肉瘤MG-63细胞的侵袭能力,作用浓度越大,侵袭力抑制作用越明显。淫羊藿素作用后的细胞中Bax蛋白表达随着淫羊藿素作用浓度的增加而逐渐降低。淫羊藿素作用后的细胞中Bcl-2、MMP-9蛋白表达随着淫羊藿素作用浓度的增加而逐渐降低。结论淫羊藿素可以抑制骨肉瘤细胞增殖和侵袭,促进骨肉瘤细胞凋亡,抑制作用随着淫羊藿素浓度的增加而增加,促细胞凋亡作用随着作用时间和作用浓度的增加而增加,作用机制与Bax、Bcl-2、MMP-9有关。

Objective To study the effect and mechanism of Icaritin on the proliferation and differentiation of osteosarcoma cells. Methods In human osteosarcoma MG-63 cells as the research object,respectively,with 4,8,16,32,64 μmol/L concentration of Icaritin for 24 h,48 h,72 h,respectivelydivided control group. Cell proliferation was detected with MTT. The cell apoptosis rate was observed and calculated by flow cytometry after Annexin V/PI double staining of MG-63 for 4,8,16,32 μmol/L of 48 h. The Transwell assay by icaritin concentration of 4,8,16,32 μmol/L role after 48 h of human osteosarcoma MG-63 cells invasion ability. Western blot assay the expression of Bax,Bcl-2 and MMP-9 in cells. Results compared with the control group,the inhibitory rate of MG-63 was significantly different from that in the control group（P 〈0. 05）. Osteosarcoma cells were treated with 0,4,8,16,32 μmol/L,apoptosis rate were 0. 125 1 ± 0. 018 4,0. 241 4 ± 0. 019 7,0. 436 5 ± 0. 030 1,0. 723 2 ± 0. 031 5,0. 789 7 ± 0. 047,further illustrates the icaritin is by promoting apoptosis of cancer cells and inhibit the proliferation of the cancer cells. The concentration of 4,8,16,32 μmol/L,after the role of osteosarcoma MG-63 cells invasion force compared with the control group significantly（P 〈0. 01）. The effect of MG-63 on the invasion of osteosarcoma cells was decreased,and the effect of the inhibition of the invasion was more obvious. The expression of Bax protein in the cells after the action of the Icaritin was decreased with the increase of the concentration. The expression of Bcl-2 and MMP-9 protein in the cells after the action of Icaritin was gradually decreased with the increase of the concentration. Conclusion Icaritin can inhibit the proliferation and invasion of osteosarcoma cells,promote apoptosis of osteosarcoma cells,the inhibition increases with the increase of icaritin concentration increased,promote cell apoptosis increased with time and concentration increase,its mechanism related to Bax,Bcl-2,MMP-9.