Hispolon from Phellinus linteus induces G0/G1 cell cycle arrest and apoptosis in NB4 human leukaemia cells.
Hispolon (a phenolic compound isolated from Phellinus linteus) has been shown to possess strong antioxidant, anti-inflammatory, anticancer, and antidiabetic properties. In this study, we investigated the antiproliferative effect of hispolon on human hepatocellular carcinoma NB4 cells using the MTT assay, DNA fragmentation, DAPI (4, 6-diamidino-2-phenylindole dihydrochloride) staining, and flow cytometric analysis. Hispolon inhibited the cellular growth of NB4 cells in a dose-dependent manner through the induction of cell cycle arrest at G0/G1 phase measured using flow cytometric analysis and apoptotic cell death, as demonstrated by DNA laddering. Exposure of NB4 cells to hispolon-induced apoptosis-related protein expressions, such as the cleavage form of caspase 3, caspase 8, caspase 9, poly (ADP ribose) polymerase, and the proapoptotic Bax protein. Western blot analysis showed that the protein levels of extrinsic apoptotic proteins (Fas and FasL), intrinsic related proteins (cytochrome c), and the ratio of Bax/Bcl-2 were increased in NB4 cells after hispolon treatment. Hispolon-induced G0/G1-phase arrest was associated with a marked decrease in the protein expression of p53, cyclins D1, and cyclins E, and cyclin-dependent kinases (CDKs) 2, and 4, with concomitant induction of p21waf1/Cip1 and p27Kip1. We conclude that hispolon induces both of extrinsic and intrinsic apoptotic pathways in NB4 human leukemia cells in vitro.
School of Post-Baccalaureate Chinese Medicine, Tzu Chi University, Hualien 970, Taiwan.
Yi-Chuan
YC
Chen
Heng-Yuan
HY
Chang
Jeng-Shyan
JS
Deng
Jian-Jung
JJ
Chen
Shyh-Shyun
SS
Huang
I-Hsin
IH
Lin
Wan-Lin
WL
Kuo
Wei
W
Chao
Guan-Jhong
GJ
Huang
D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics*; D002396:Catechols / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use*; D059447:Cell Cycle Checkpoints / Q000187:drug effects* / Q000235:genetics*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D052138:Genes, Neoplasm / Q000235:genetics; D006801:Humans; D007938:Leukemia / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology* / Q000517:prevention & control; D000084682:Phellinus; D008517:Phytotherapy; D010936:Plant Extracts; D011134:Polysaccharides / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use*
D000972:Antineoplastic Agents, Phytogenic; D002396:Catechols; C526145:Phellinus linteus extract; D010936:Plant Extracts; D011134:Polysaccharides; C103442:hispolon