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姜黃素對人骨肉瘤細(xì)胞的生長抑制及對MAPK基因的調(diào)控作用研究

發(fā)布時(shí)間:2019-01-15 23:27
【摘要】:目的:研究姜黃素人成骨肉瘤細(xì)胞MG-63的生長抑制作用,并檢測對骨肉瘤細(xì)胞中MAPK基因表達(dá)的調(diào)控作用,初步明確姜黃素對骨肉瘤細(xì)胞的作用機(jī)制。方法:以人骨肉瘤細(xì)胞株MG-63為研究對象,采用不同劑量姜黃素處理。MTT法檢測5,10,15,20,25,30,35,40,50μmol/L姜黃素作用24,48,72 h對細(xì)胞的毒性作用;Transwell小室侵襲實(shí)驗(yàn)法檢測5,10,15μmol/L姜黃素對細(xì)胞的侵襲能力,黏附實(shí)驗(yàn)法檢測對細(xì)胞的黏附能力;Western blot法檢測5,10,15μmol/L姜黃素作用后MAPK蛋白表達(dá)水平的影響,RT-PCR法檢測對細(xì)胞中MAPK基因m RNA轉(zhuǎn)錄水平的影響。結(jié)果:姜黃素濃度15μmol/L,作用時(shí)間24 h時(shí),對細(xì)胞的生長抑制呈時(shí)間和劑量依懶性(P0.001),當(dāng)姜黃素濃度≤15μmol/L,作用時(shí)間為24 h時(shí),對細(xì)胞無明顯毒性作用,細(xì)胞存活率85%;細(xì)胞侵襲能力和黏附能力隨著姜黃素濃度的增加而逐漸降低,以15μmol/L姜黃素濃度處理效果最明顯(P0.05);姜黃素可明顯抑制MAPK基因m RNA轉(zhuǎn)錄水平,其抑制作用與劑量呈高度依懶性(P0.05)。結(jié)論:姜黃素可通過抑制MAPK信號通路的活化及此信號通路靶基因MAPK的表達(dá)降低人骨肉瘤細(xì)胞株MG-63的侵襲性。
[Abstract]:Aim: to investigate the inhibitory effect of curcumin on the growth of human osteosarcoma cell line MG-63, and to investigate the effect of curcumin on the expression of MAPK gene in osteosarcoma cells, and to clarify the mechanism of curcumin on osteosarcoma cells. Methods: human osteosarcoma cell line MG-63 was treated with different doses of curcumin. The cytotoxicity of curcumin on human osteosarcoma cell line (MG-63) was detected by MTT method. The invasiveness of curcumin to cells was detected by Transwell chamber invasion assay, and the adhesion ability of curcumin to cells was detected by adhesion assay. Western blot assay was used to detect the effect of curcumin on the expression of MAPK protein, and RT-PCR assay was used to detect the transcription level of MAPK gene m RNA. Results: when the concentration of curcumin was 15 渭 mol/L, for 24 h, the inhibition of cell growth was time-dependent and dose-dependent (P0.001). When the concentration of curcumin was 鈮,

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