Lu Jin-Jian, Lu De-Zhao, Chen Yu-Fei, Dong Ya-Ting, Zhang Jun-Ren, Li Ting, Tang Zheng-Hai, Yang Zhen
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.
College of Life Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Chin J Nat Med. 2015 Sep;13(9):673-9. doi: 10.1016/S1875-5364(15)30065-0.
Platycodin D (PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins (i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.
桔梗皂苷D(PD)是从桔梗中分离出的一种三萜皂苷,桔梗是一种著名的中药材,已被证明在多种癌细胞系中具有抗增殖作用。本研究的目的是使用蛋白质组学方法确定用PD处理肝癌HepG2细胞后细胞蛋白质的变化。使用MTT法测定细胞活力。通过二维差异凝胶电泳和基质辅助激光解吸/电离飞行时间质谱分析蛋白质组。蛋白质免疫印迹分析用于确认变化蛋白质的表达。我们的结果表明,PD以浓度和时间依赖性方式抑制HepG2细胞的增殖。在经PD处理的HepG2细胞中,有16种蛋白质被鉴定为上调,包括ATP5H、OXCT1、KRT9、CCDC40、ERP29、RCN1、ZNF175、HNRNPH1、HSP27、PA2G4、PHB、BANF1、TPM3、ECH1、LGALS1和MYL6。用PD处理后,HepG2细胞中的三种蛋白质(即RPS12、EMG1和KRT1)减少。蛋白质免疫印迹进一步证实了HSP27和PHB的变化。总之,我们的结果为PD抗癌活性的作用机制提供了新的线索。