穿心莲内酯对人骨肉瘤细胞的影响及其分子机制  

EFFECT OF ANDROGRAPHOLIDE ON HUMAN OSTEOSARCOMA CELLS AND ITS MOLECULAR MECHANISM

作　　者：张希劲 陈涛[2] 周缜[3] 沈若武[1] 刘小强 李兆 杨希重[5] ZHANG Xijin;CHEN Tao;ZHOU Zhen;SHEN Ruowu;LIU Xiaoqiang;LI Zhao;YANG Xizhong(Department of Special Medicine, School of Basic Medicine, Qingdao University, Qingdao 266071, China)

机构地区：[1]青岛大学基础医学院特种医学系,山东青岛266071 [2]青岛大学附属医院放射科 [3]青岛大学外科实验中心 [4]青岛大学医学部 [5]即墨市人民医院骨科

出　　处：《精准医学杂志》2020年第2期100-105,共6页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81274116);山东省高等学校科技计划项目(L16LK04)。

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、100μmol/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=9608.52、5409.49,F时间=8312.14、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。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 100μmol/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=9608.52,5409.49,Ftime=8312.14,352.91,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 ki-lling effect on human osteosarcoma cells,possibly by affecting the SphK1 signaling pathway.

关 键 词：穿心莲内酯 骨肉瘤 细胞系 肿瘤 细胞增殖 细胞运动 肿瘤侵润 细胞周期 鞘氨醇激酶 黏着斑蛋白酪氨酸激酶类 

分 类 号：R285.5[医药卫生—中药学] R738.1[医药卫生—中医学]