Abstract
Background Paris polyphylla is an oriental folk medicine that has anticancer activities both in vivo and in vitro. Polyphyllin VII (PP7), a pennogenyl saponin from P. polyphylla has been found to exert strong anticancer activity. However, the underlying mechanisms are poorly understood. In the present study, the anticancer effect of polyphyllin VII against human liver cancer cells and the molecular mechanisms were investigated.MethodsCellular viability was measured by MTT assay. Apoptosis, intracellular reactive oxygen species (ROS) and mitochondrial membrane potential levels were evaluated using the InCell 2000 confocal microscope. The expression levels of apoptotic-related proteins were evaluated by Western blotting.ResultsPP7 strongly inhibited the cell growth and induced apoptosis and necrosis in hepatocellular carcinoma HepG2 cells. Meanwhile, PP7 up-regulated the levels of Bax/Bcl-2, cytochrome c, the cleaved forms of caspases-3, -8, -9, and poly (ADP-ribose) polymerase in a dose- and time-dependent manner, indicating that PP7 induced apoptosis in HepG2 cells through both intrinsic and extrinsic pathways. Moreover, PP7 provoked the production of intracellular ROS and the depolarization of mitochondrial membrane potential. Further analysis showed that PP7 significantly augmented the phosphorylation of JNK, ERK and p38, the major components of mitogen-activated protein kinase (MAPK) pathways, and the expressions of tumor suppressor proteins p53 and PTEN. In addition, PP7-induced apoptosis was remarkably attenuated by MAPK inhibitors and ROS inhibitor.ConclusionsThese results demonstrated that PP7 induced apoptotic cell death in HepG2 cells through both intrinsic and extrinsic pathways by promoting the generation of mitochondrial-mediated ROS and activating MAPK and PTEN/p53 pathways.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1036-x) contains supplementary material, which is available to authorized users.
Highlights
Paris polyphylla is an oriental folk medicine that has anticancer activities both in vivo and in vitro
Primary antibodies against Bcl-2, Bax, cytochrome c, p-Bcl2-associated agonist of cell death (BAD), BAD, p-stat3, stat3, p-phosphatase and tensin homolog (PTEN), PTEN, extracellular signal-regulated kinase (ERK), p-ERK, p38 mitogen-activated protein kinase (MAPK) (p38), p-p38, c-Jun N-terminal kinase (JNK), p-JNK, p53, cleaved caspase-3, -8, -9 and poly (ADP-ribose) polymerase (PARP) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and secondly antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) or Proteintech Group Inc (Wuhan, Hubei, China)
The results showed that polyphyllin VII (PP7) significantly inhibited the growth of all cell lines, with 50 % inhibitory concentration (IC50) values in the range from 1.32 to 2.70 μM
Summary
Paris polyphylla is an oriental folk medicine that has anticancer activities both in vivo and in vitro. The anticancer effect of polyphyllin VII against human liver cancer cells and the molecular mechanisms were investigated. Numerous studies have demonstrated that apoptosis can be triggered by diverse pro-apoptosis stimuli converging on mitochondria, causing mitochondrial depolarization, cytochrome c release, caspase enzymes activation and eventually apoptotic cell death [8]. PTEN is one of the most frequently mutated, deleted, or silenced tumor suppressor genes in cancers. It regulates p53 transcriptional activity and protein levels [9]. Many findings have demonstrated that PTEN and p53 played a critical role in DNA damage response and regulated cell cycle and apoptosis [11, 12]
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