DIM增强去甲氧柔红霉素对人前列腺癌细胞生长的抑制作用
3,3-diindolyimethane enhances the inhibitory effect of idarubicin on the growth of human prostate cancer cells
目的 观察3,3-二吲哚基甲烷(DIM)与去甲氧柔红霉素(IDA)联合应用对人前列腺癌细胞(PC-3M)的生长抑制的作用并探讨其机制.方法 应用MTT法检测生长抑制率;流式细胞术及吖啶橙染色分析PC-3M细胞凋亡及细胞周期变化;分别用RT-PCR和Western印迹检测凋亡相关的半胱氨酸蛋白水解酶9(caspase 9)基因和蛋白的表达.结果 0.5 mg/L的IDA与60 μmol/L的DIM联用,使0.5 mg/L的IDA对肿瘤细胞生长抑制率由27.8%提高到69.9%;诱导凋亡的效应由3.2%提高到47.0%,相当10倍剂量(5 mg/L)IDA产生的效应.RT-PCR和Western印迹结果显示,两药联用明显增强caspase 9基因和蛋白的表达水平.结论 DIM能显著增强IDA对PC-3M细胞的生长抑制作用,其机制与凋亡诱导效应有关.
更多Objective To study the effects of idarubicin(IDA)combined with 3,3-diindolylmethane(DIM)on the growth inhibition of human prostate cancer cells. Methods Human prostate cancer cells of the line PC-3M were cultured and then divided into the following groups: control group with solvent added into the culture fluid: IDA groups, with IDA of the terminal concentrations of 0.5,1 or 5 mg/L added into the culture fluid; DIM groups, with DIM of the terminal concentrations of 30,60 or 100 μmol/L added into the culture fluid; and DIM+IDA groups, with 0.5 mg/L IDA and DIM 30,60 or 100 μmol/L added into the culture fluid.48 h later the cell growth inhibition rate was detected by MTT assay. Flow cytometry and acridine orange staining were used to detect the cell cycle and apoptosis. RT-PCR and Western blotting were used to detect the mRNA and protein expression of caspase 9,an apoptosis gene. Results Both IDA and DIM dose-dependently inhibited the growth of the PC-3M cells. The growth inhibition rate of the 60 μmol/L DIM+0.5 mg/L IDA group was 69.9%,almost 10 times as that of the 0.5 mg/L IDA group. The apoptosis rate of the 60 μmol/L DIM+0.5 mg/L IDA group was 47.0%,significantly higher than that of the 0.5 mg/L IDA group(3.2%,P<0.05).RT-PCR and Western blotting showed- that the combination of DIM and IDA significantly enhanced the mRNA and protein expression of caspase 9. Conclusion DIM enhances the growth inhibition effect of IDA on human prostate cancer cells by the mechanism of induction of apoptosis.
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