DIM联合顺铂诱导PC-3细胞凋亡机制的研究

The apoptosis mechanism of DDP associating with DIM inducing Human Prostate Cancer Cell PC-3

目的探讨顺铂(DDP)和3,3-二吲哚基甲烷(3,3-diindolylmethane,DIM)联合诱导人前列腺癌PC-3细胞凋亡的机制。方法采用免疫组化技术观察细胞内bcl-2,caspase3和caspase9蛋白的表达,利用RT-PCR和western blot检测bcl-2,caspase3和caspase9基因和蛋白表达变化。结果细胞培养及免疫组化可见:各给药组与对照组比较均有不同程度抑制PC-3细胞增殖并诱导其凋亡作用;RT-PCR结果显示:各给药组均能使caspase3和caspase9基因表达上调,bcl-2表达下调;western blot结果表明:各给药组与对照组相比caspase3和caspase9蛋白表达上调,bcl-2表达下调,且0.4mg.L-1DDP+60μmol.L-1DIM联合应用与10倍剂量DDP(4 mgL-1)单独应用效果相同。结论DIM与DDP均可抑制PC-3细胞增殖并诱导其凋亡;其细胞凋亡机制可能与上调caspase3、caspase9基因表达,下调bcl-2表达有关;DIM与DDP联合抗瘤具有明显的减毒增效作用。

Objective To study the apoptesis mechanism of 3, 3-diindolylmethane （DIM） associating with diamminedichloreplatinum（DDP） on human prostate cancer cell PC-3. Methods The expressions of the bcl-2, easpase3 and caspase9 in the cells by immunotfistochemistry, at the same time the expressions of bcl-2, easpase3 and caspase9 were detected by RT-PCR and western blotting. Results Compared with controlled group, the proliferation of PC-3 ceils was inhibited and the apoptosis was induced differently in all treatment groups through immunohistochemistry and cell culture;The RT-PCR and western blotting results manifested that the expressions of caspase3 and caspase9 were upregulated but Bcl-2 was downregulated in treatment groups compared with the controlled group. Combining 60μmol.L^-1 DIM with 0.4mg.L^-1 DDP could effectively inhibit the growth of PC-3 cell. This experiment result is the same as the single work of 4 mg.L^-1 DDP. Conclusion Both DIM and DDP can inhibit PC-3 cell proliferation and induce cell apoptosis. The mechanism may be that the both upregulate the expression of caspase3 and easpase9 but downregulate Bel- 2. DIM associating with DDP has an obvious role in attenuated toxcity and Synergies.