摘要
目的:研究姜黄素人成骨肉瘤细胞MG-63的生长抑制作用,并检测对骨肉瘤细胞中MAPK基因表达的调控作用,初步明确姜黄素对骨肉瘤细胞的作用机制。方法:以人骨肉瘤细胞株MG-63为研究对象,采用不同剂量姜黄素处理。MTT法检测5,10,15,20,25,30,35,40,50μmol/L姜黄素作用24,48,72 h对细胞的毒性作用;Transwell小室侵袭实验法检测5,10,15μmol/L姜黄素对细胞的侵袭能力,黏附实验法检测对细胞的黏附能力;Western blot法检测5,10,15μmol/L姜黄素作用后MAPK蛋白表达水平的影响,RT-PCR法检测对细胞中MAPK基因m RNA转录水平的影响。结果:姜黄素浓度>15μmol/L,作用时间>24 h时,对细胞的生长抑制呈时间和剂量依懒性(P<0.001),当姜黄素浓度≤15μmol/L,作用时间为24 h时,对细胞无明显毒性作用,细胞存活率>85%;细胞侵袭能力和黏附能力随着姜黄素浓度的增加而逐渐降低,以15μmol/L姜黄素浓度处理效果最明显(P<0.05);姜黄素可明显抑制MAPK基因m RNA转录水平,其抑制作用与剂量呈高度依懒性(P<0.05)。结论:姜黄素可通过抑制MAPK信号通路的活化及此信号通路靶基因MAPK的表达降低人骨肉瘤细胞株MG-63的侵袭性。
Objective：To investigate the regulatory effect of curcumin on expression of MAPK in human osteosarcoma as well as the possible mechanism of curcumin in prevention and treatment of human osteosarcoma. Methods ： Highly invasive MG -63 cells were treated with curcumin at various dosages. The cytotoxic effects of treatment with 5 μmol/L, 10 μmol/L, 15 μmol/L,20 μmol/L,25 μmol/L,30 μmol/L,35 μmol/L,40 μmol/L and 50 μmol/L curcumin for 24 h,48 h and 72 h on MG - 63 cells were measured by MTT assay. The invasion abilities of cells treated with 5 μmol/L, 10 μmol/L and 15 μmol/L curcumin were measured by Transwell test while the adhesive abilities by cell adhesion assay. The effect of 5μmol/L, 10 μmol/L and 15 μmol/L curcumin on expression levels of MAPK were determined by western blot while those on transcription level of MAPKmRNA by RT - PCR. Results：The treatment with curcumin at a dosage more than 15 μmol/L for more than 24 h inhibited the growth of MG -63 cells in time - and dose - dependent patterns（ P 〈 0. 001 ）. The treatment with eurcumin at a dosage not less than 15 μmol/L for 24 h showed no significant eytotoxic effect on the ceils while the survival rate of cells was more than 85%. The invasion and adhesive abilities of cells were decreased gradually with the increasing eureumin concentration. The eureumin at a concentration of 15 μmol/L showed the strongest inhibitory effect on the two abilities （ P 〈 0. 05 ）. Curcumin showed significant dose - dependent inhibitory effect on the transcription level of MAPK mRNA（ P 〈 0. 05 ）. Conclusions： Curcumin inhibited the invasive abilitiy of MG - 63 cells by inhibiting the activation of MAPK signal pathway and expression of MAPK was as a target gene of the path- way.
出处
《中华中医药学刊》
CAS
北大核心
2017年第2期428-431,I0017,共5页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省中医药科技计划项目(2015ZB054)
