摘要
目的研究兔儿风属植物中天然产物gochnatiolide A抗前列腺癌DU145细胞的作用及机制。方法采用CCK-8法和克隆形成实验检测gochnatiolideA对DU145细胞增殖的影响;DAPI染色法观察细胞凋亡形态;流式细胞仪检测gochnatiolide A对DU145细胞凋亡和周期的影响;Western blotting法检测gochnatiolide A对DU145细胞内活化的多聚腺苷二磷酸核糖聚合酶(cleaved-PARP)、活化的半胱氨酸蛋白酶9(cleaved Caspase-9)、p53、Bcl-2、核转录因子κB p65(NF-κB p65)和核转录因子κB抑制蛋白α、β(IKKα、IKKβ)表达的影响。结果与对照组比较,gochnatiolideA能够显著抑制DU145细胞增殖,其12、24、48 h的半数抑制浓度(IC_(50))值分别为4.15、2.80、1.74μmol/L。Gochnatiolide A可以浓度依赖性地促进细胞凋亡,阻滞细胞周期于G2期和S期;可以显著促进凋亡蛋白cleaved-PARP、cleaved Caspase-9和p53蛋白表达,抑制Bcl-2蛋白表达,抑制NF-κB通路中NF-κB p65、IKKα和IKKβ蛋白的表达。结论 Gochnatiolide A具有很好的抑制DU145细胞增殖及促进凋亡的作用,其机制可能与抑制NF-κB通路有关。
Objective To investigate the effects and mechanism of gochnatiolide A from Ainsliaea in anti-prostate cancer. Methods The activities of gochnatiolide A on the proliferation of DU145 cells were tested by CCK-8 and colony formation experiments.Apoptosis morphological changes of cells were observed by DAPI staining. The apoptosis and cell cycle were evaluated by flow cytometry. The expressions of cleaved Poly ADP ribose polymerase(cleaved-PARP), cleaved cystein-containing aspartate specific protease 9(cleaved Caspase-9), tumor suppressor protein(p53), b cell lymphoma-2(Bcl-2), transcription factors-κB p65(NF-κB p65)and IκB kinase α, β(IKKα, IKKβ) proteins in DU145 cells were investigated by Western blotting. Results The natural product gochnatiolide A significantly inhibited the proliferation of prostate cancer DU145 cells, with IC_(50) values of 4.15, 2.80 and 1.74 μmol/L at 12 h, 24 h, and 48 h, respectively. Meanwhile, gochnatiolide A promoted DU145 cells apoptosis and induced cell cycle arrest at G2 and S phases in a concentration dependent manner. In addition, gochnatiolide A also up-regulated apoptosis proteins cleaved-PARP,cleaved Caspase-9 and p53 proteins, and down-regulated Bcl-2 protein. Comparing the control group, the expression of NF-κB p65, IKKαand IKKβ proteins in the NF-κB pathway were decreased after treatment with gochnatiolide A. Conclusion The natural product gochnatiolide A remarkably inhibited the proliferation and promoted apoptosis in DU145 cells, and the possible mechanism was related to the inhibition of NF-κB pathway.
引文
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