Daphnoretin Induces Cell Cycle Arrest and Apoptosis in Human Osteosarcoma (HOS) Cells

• Published in: Molecules (Basel, Switzerland) Vol. 17; no. 1; pp. 598 - 612
• Main Authors: Gu, Shoubin, He, Jinhai
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 09.01.2012; MDPI
• Subjects: Antineoplastic Agents, Phytogenic - pharmacology; antiproliferative; Apoptosis; Apoptosis - drug effects; Apoptosis Regulatory Proteins - genetics; Apoptosis Regulatory Proteins - metabolism; Bone cancer; CDC2 Protein Kinase; Cell cycle; cell cycle arrest; Cell death; Cell growth; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Shape - drug effects; Coumarins - pharmacology; Cyclin A - metabolism; Cyclin B - metabolism; Cyclin B1 - metabolism; Cyclin-Dependent Kinases; Cytochrome; Cytochromes c - metabolism; Cytotoxicity; daphnoretin; Drug dosages; G2 Phase Cell Cycle Checkpoints - drug effects; Gene Expression; HOS; Humans; Inhibitory Concentration 50; Kinases; Membrane Potential, Mitochondrial - drug effects; Osteosarcoma; Telomerase; Tumors; China
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules17010598

In this study antiproliferation, cell cycle arrest and apoptosis induced by daphnoretin in human osteosarcoma (HOS) cells were investigated. Antiproliferative activity was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The IC50 value of daphnoretin was 3.89 μM after 72 h treatment. Induction of apoptosis was evidenced by apoptotic body appearance and Annexin V-FITC/PI apoptosis detection kit. Flow cytometric analysis indicated daphnoretin arrested the cell cycle in the G2/M phase. Western-blot assay showed that the G2/M phase arrest was accompanied by down-regulation of cdc2, cyclin A and cyclin B1. Moreover, daphnoretin inhibited Bcl-2 expression and induced Bax expression to desintegrate the outer mitochondrial membrane and causing cytochrome c release. Mitochondrial cytochrome c release was associated with the activation of caspase-9 and caspase-3 cascade. Our results demonstrated that daphnoretin caused death of HOS cells by blocking cells successively in G2/M phases and activating the caspase-3 pathway.