白花丹素体外诱导前列腺癌PC-3细胞凋亡的研究

Apoptosis of human prostate cancer cell line PC-3 induced by plumbagin

目的 探讨白花丹素对前列腺癌PC-3细胞增殖、凋亡的作用及其和RelA（p65）表达的关系.方法 应用不同浓度梯度的白花丹素（1、5、1O、15、20 μmol/L）共同培养PC-3细胞24、48 h,噻唑蓝（MTT）比色法检测PC-3细胞增殖活力,双染流式细胞仪检测凋亡细胞,透射电镜观察超微病理变化,计算药物半数抑制浓度（IC50）.逆转录-聚合酶链反应（RT-PCR）法扩增检测RelA（p65）.结果 24 h组在10～20 μmol/L,48 h组在5～20 μmol/L时均出现生长抑制,IC50分别为12.88、3.71 μmol/L.双染流式细胞仪检测显示PC-3随作用浓度的上升凋亡率增加并呈现浓度依赖关系.透射电镜观察作用后PC-3呈现典型凋亡表现.RT-PCR结果提示其细胞凋亡率同Rel A（p65）表达呈负相关.结论 白花丹素体外实验可能通过抑制Rel A（p65）表达诱导PC-3的凋亡.

Objective To study the effect of human prostate cancer cell line PC-3 apoptosis induced by plumbagin in vitro, and the relation to the expression of Rel A（ p65 ）. Methods PC-3 cells were co-cultured with the desired concentrations of plumbagin （1, 5, 10, 15, 20μjnol/L） for 24, 48 h. Cell inhibitory rate was measured by MTT colorimetric assay. Apoptosis-inducing effect was determined by double stained flow cytometry. The cell apoptosis morphologic changes were observed by a transmission e-lectron microscope. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the expression of Rel A（ p65）. Results The growth inhibition of PC-3 cells appeared in 5 to 20μmol/L plumbagin solution for 24 h and 10 to 20 μmol/L for 48 h. IC50 of 24 h was 13.88μmol/L and 3.71μmol/L for 48 h, respectively. The apoptosis and the death rate of the cells showed that plumbagin reduced PC-3 cells apoptosis in a dose-dependent manner between 5 and 20μmol/L. RT-PCR revealed that the expression of p65 was reversely correlated with the apoptosis rate. Conclusion Plumbagin may inhibit human prostate cancer cell PC-3 proliferation and induce apoptosis of PC-3 cell line by suppressing p65 in vitro.