双氢青蒿素对人骨巨细胞瘤细胞活性和迁移能力的影响

Effects of dihydroartemisinin on migration and cell viability of human giant cell tumor cells of bone

目的 观察双氢青蒿素（DHA）对人骨巨细胞瘤细胞活性、凋亡和迁移能力的影响。 方法 将不同浓度的DHA（0、10、20 μmol/L）作用于人骨巨细胞瘤细胞24 h后，膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）/碘化丙锭（PI）染色，流式细胞仪检测细胞的凋亡；噻唑蓝（MTT）法检测人骨巨细胞瘤细胞活性；Transwell实验检测瘤细胞的迁移和侵袭能力；同时用Western blot检测瘤细胞中基质金属蛋白酶（MMP）-2、MMP-9蛋白的表达。结果 DHA作用24 h后，0、10、20 μmol/L组人骨巨细胞瘤细胞凋亡率分别为0.16%、18.57%和34.28%，差异有统计学意义（P＝0.000、0.000）。10、20 μmol/L DHA组细胞的相对活性分别为（60.63±2.45）%和（45.93±9.36）%，DHA作用组细胞活性明显低于对照组（P＝0.024、0.000）；DHA作用组细胞的相对迁移能力[分别为（69.27±4.82）%和（46.43±3.96）%]和侵袭能力[分别为（61.98±9.37）%和（43.55±2.45）%]均明显低于对照组细胞。Western blot结果显示，与对照组比较，10、20 μmol/L DHA组MMP-2（分别为0.37±0.05和0.13±0.01）和MMP-9（分别为0.48±0.01和0.23±0.00）的相对表达量均明显降低。结论 DHA能够抑制骨巨细胞瘤细胞活性、促进瘤细胞凋亡，同时可通过抑制MMP-2和MMP-9的表达而限制骨巨细胞瘤的迁移和侵袭。

Objective To research the effects of dihydroartemisinin （DHA） on migration and cell viability of human giant cell tumor cells of bone. Methods The human giant cell tumor cells of bone were treated with 0, 10, 20, 40 μmol/L DHA for 24 h. After treated, the cells were stained with Annexin V-fluoresceine isothiocyanate （FITC）/propidium iodide （PI） and the apoptosis rate was measured by flow cytometry. At the same time the mitochondrial membrane potential change was measured.Results After treated with DHA for 24 h, the apoptosis rates of human giant cell tumor cells of bone （0, 10, 20 μmol/L DHA） were respectively 0.16%, 18.57% and 34.28%. Compared with control group, the apoptosis rates of human giant cell tumor cells of bone in DHA groups were significantly decreased （P=0.000, 0.000）. The relative cell viability were respectively （60.63±2.45）% and （45.93±9.36）% in 10 and 20 μmol/L DHA group, which were significantly decreased than that in control group （P=0.024, 0.000）. The migration [（69.27±4.82）% and （46.43±3.96）%] and invasion [（61.98±9.37）% and （43.55±2.45）%] of human giant cell tumor cells of bone in 10 and 20 μmol/L DHA group, which were significantly decreased than those in control group. Compared with control group, the expression levels of matrix metalloproteinase （MMP）-2 and MMP-9 were decreased.Conclusion DHA can significantly inhibit the cell viability and increase cell apoptosis. At the same time, DHA can inhibit the migration and invasion of human giant cell tumor cells of bone through decreasing the expression levels of MMP-2 and MMP-9.