%0 Journal Article %T 穿心莲内酯对人骨肉瘤细胞的影响及其分子机制 %A 张希劲　陈涛　周缜　沈若武　刘小强　李兆　杨希重 %J - %D 2020 %R 10.13362/j.jpmed.202002002 %X 摘要目的　探讨穿心莲内酯(AG)对人骨肉瘤细胞的影响及其分子机制。方法　分别以含有AG为0、5、10、20、50、100 μmol/L（A、B、C、D、E、F组）的培养基培养人骨肉瘤细胞，倒置显微镜下计数各组细胞数，并计算各组细胞密度及细胞生存率；采用平板克隆实验观察AG对人骨肉瘤HOS和U2OS细胞增殖能力的影响；Transwell小室实验观察AG对人骨肉瘤HOS和U2OS细胞侵袭和迁移能力的影响；流式细胞术观察AG对人骨肉瘤HOS和U2OS细胞周期分布的影响；Western Blot实验检测AG对鞘氨醇激酶（SphK1）以及黏着斑激酶（FAK）表达水平的影响。结果　组别、时间以及组别与时间交互作用均对人骨肉瘤HOS和U2OS细胞的生存率具有明显影响（F组别=9 608.52、5 409.49，F时间=8 312.14、352.91，F时间*组别=18.06、3.55，P＜0.05）；使增殖能力呈浓度依赖性降低（FHOS=97.41，FU2OS=174.13，P＜0.05）；侵袭和迁移能力明显减弱（t=39.06～135.96，P＜0.05）；组别、周期及组别与周期交互作用均对人骨肉瘤HOS和U2OS细胞的细胞周期分布具有明显影响（F组别=35.05、8.24，F周期=3211.19、192.49，F周期*组别=75.96、140.97，P＜0.05）；SphK1和FAK的表达水平均下调（t=15.94～95.79，P＜0.05）。结论　AG对人骨肉瘤细胞有杀伤作用，其可能与AG影响SphK1信号通路有关。
Abstract：Objective　To investigate the effect of andrographolide (AG) on human osteosarcoma cells and its molecular mechanism. Methods　Human osteosarcoma cells were cultured in a medium containing AG at a concentration of 0, 5, 10, 20, 50, and 100 μmol/L (groups A, B, C, D, E, and F, respectively). An inverted microscope was used to measure the number of cells and calculate cell density and viability in each group; plate colony formation assay was used to observe the effect of AG on the proliferative capacity of human osteosarcoma HOS and U2OS cells; Transwell chamber assay was used to observe the effect of AG on the invasion and migration abilities of human osteosarcoma HOS and U2OS cells; flow cytometry was used to observe the effect of AG on cell cycle distribution of human osteosarcoma HOS and U2OS cells; Western Blot was used to observe the effect of AG on the expression levels of sphingosine kinase 1 (SphK1) and focal adhesion kinase (FAK). Results　Group, time, and group-time interaction had significant influence on the viability of human osteosarcoma HOS and U2OS cells (Fgroup=9 608.52,5 409.49,Ftime=8 312.14,352.91,Ftime*group=18.06,3.55,P<0.05). The proliferation capacity decreased in a concentration-dependent manner (FHOS=97.41,FU2OS=174.13,P<0.05), and there were significant reductions in the invasion and migration abilities (t=39.06-135.96,P<0.05). Group, period, and group-period interaction had significant influence on the cell cycle distribution of human osteosarcoma HOS and U2OS cells (Fgroup=35.05,8.24,Fperiod=3211.19,192.49,Fgroup*period=75.96,140.97,P<0.05). The expression levels of SphK1 and FAK were significantly downregulated (t=15.94-95.79,P<0.05). Conclusion　AG has a killing effect on human osteosarcoma cells, possibly by affecting the SphK1 signaling pathway %K 穿心莲内酯 %K 骨肉瘤 %K 细胞系 %K 肿瘤 %K 细胞增殖 %K 细胞运动 %K 肿瘤侵润 %K 细胞周期 %K 鞘氨醇激酶 %K 黏着斑蛋白酪氨酸激酶类
Andrographolide %K Osteosarcoma %K Cell line %K tumor %K Cell proliferation %K Cell movement %K Neoplasm invasiveness %K Cell cycle %K Sphingosine kinase %K Focal adhesion protein-tyrosine kinases %U http://jpmed.qdu.edu.cn/CN/10.13362/j.jpmed.202002002