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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 study the inhibitory effect of curcumin on human osteosarcoma cell line MG-63 and its role in the regulation of MAPK gene expression in osteosarcoma cells, and to clarify the mechanism of curcumin on osteosarcoma cells. Methods: The human osteosarcoma cell line MG-63 as the research object, using different doses of curcumin treatment. MTT assay was used to detect the toxic effects of curcumin at 5, 10, 15, 20, 25, 30, 35, 40 and 50μmol / L for 24,48,72 h; Transwell chamber invasion assay was used to detect 5,10,15μmol / The effect of curcumin on the invasion of cells and the adhesion ability of cells were detected by adhesion assay. The effect of curcumin at 5, 10 and 15μmol / L on MAPK protein expression was detected by Western blot. The effect of MAPK gene m RNA transcriptional level. RESULTS: Curcumin concentration> 15 μmol / L for 24 h resulted in time-and dose-dependent inhibition of cell growth (P <0.001). When curcumin concentration was ≤15 μmol / L for 24 h , No obvious cytotoxic effect on cells, cell survival rate> 85%; cell invasiveness and adhesion decreased with the increase of curcumin concentration, 15μmol / L curcumin concentration treatment effect most obvious (P <0.05) Suprostly inhibited the transcription level of m RNA of MAPK gene. The inhibitory effect was dose-dependently lazy (P <0.05). Conclusion: Curcumin can reduce the invasiveness of human osteosarcoma cell line MG-63 by inhibiting the activation of MAPK signaling pathway and the expression of MAPK, a target gene of this signaling pathway.