[Apoptosis induced by curcumin and its effect on c-myc and caspase-3 expressions in human melanoma A375 cell line].

Shi Qiu, Sheng-shun Tan, Jiang-an Zhang, An Liu, Jing-yi Yuan, Guo-zhou Rao, Wen-yong Wang
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引用次数: 0

Abstract

Objective: To investigate the effect of curcumin on cell apoptosis and c-myc and caspase-3 expressions in human melanoma A375 cell line.

Methods: A375 cells were exposed to curcumin treatment and growth inhibition of the cells was examined by MTT assay. Annexin V/propidium iodide double staining and DNA fragmentation analysis were employed for assay of the cell apoptosis and morphological changes of the cells were observed with inverted microscopy and transmission electron microscopy, respectively. In situ hybridization and SABC immunohistochemistry were performed for detection of the expressions of c-Myc and caspase-3 in the A375 cells.

Results: Curcumin inhibited the growth of A375 cells in both time- and concentration-dependent manners. After treatment with 30 micromol/L curcumin for 48 h, apoptotic morphological changes were observed in the cells and an oligonucleosomal DNA ladder was clearly visualized in DNA fragmentation analysis. The apoptotic rates of the cells treated with curcumin at the concentration above 20 micromol/L were significantly higher than that of the control cells. c-myc expression level was decreased whereas caspase-3 expression increased with the increase in curcumin concentrations.

Conclusion: Curcumin can inhibit the proliferation and induce apoptosis of A375 cells in vitro, and the genes encoding c-myc and caspase-3 may play a role in the process.

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姜黄素诱导人黑色素瘤A375细胞株凋亡及其对c-myc和caspase-3表达的影响
目的:探讨姜黄素对人黑色素瘤A375细胞株细胞凋亡及c-myc和caspase-3表达的影响。方法:采用MTT法检测姜黄素对A375细胞的生长抑制作用。采用膜联蛋白V/碘化丙啶双染色法和DNA片段化法检测细胞凋亡情况,倒置显微镜和透射电镜观察细胞形态学变化。采用原位杂交和SABC免疫组化方法检测c-Myc和caspase-3在A375细胞中的表达。结果:姜黄素对A375细胞的生长具有时间依赖性和浓度依赖性。30微mol/L姜黄素作用48 h后,观察到细胞凋亡形态学改变,DNA片段化分析清晰可见寡核体DNA阶梯。姜黄素浓度大于20微mol/L时,细胞凋亡率显著高于对照细胞。随着姜黄素浓度的升高,C-myc表达水平降低,caspase-3表达水平升高。结论:姜黄素对体外培养的A375细胞具有抑制增殖和诱导凋亡的作用,其编码基因c-myc和caspase-3可能参与了这一过程。
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