姜黄素诱导人黑色素瘤A375细胞凋亡以及对c-myc、caspase-3表达的影响

目的 探讨姜黄素诱导人黑色素瘤A375细胞凋亡及其对C-myc、Caspase-3表达的影响。方法 姜黄素处理A375细胞，MTT法检测细胞的牛长活性，应用倒置显微镜和透射电镜进行形态学观察．采用Annexin V／PI双染法和DAN片段化分析检测细胞凋亡，SABC免疫组化法和原位杂交检测细胞内c-myc、Caspase-3表达水平：结果姜黄素能抑制A375细胞增殖，并呈剂量和时间依赖性：经30μmol／L姜黄素处理A375细胞48h，细胞旱现凋亡形态学改变。DAN片段化分析可见到明显的DNA梯带形成；流式细胞仪定量检测出20μmol／L以上浓度姜黄素诱导细胞凋亡的发生率较对照组有显著性差异。c-myc表达水平减少，而Caspase-3表达增加。结论 姜黄素能抑制A375细胞增殖和诱导其凋亡，c-myc和Caspase一3基因可能参与凋亡过程。

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

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.