Induction of apoptosis by diphenyldifluoroketone in osteogenic sarcoma cells is associated with activation of caspases.

Abstract

The aim of the present study was to investigate and compare the effects of diferuloylmethane (curcumin) and diphenyldifluoroketone (EF-24) on cell growth and apoptosis induction in human osteogenic sarcoma cells. This was examined by MTT assay, nuclear DAPI staining, caspase-activation assay, flow cytometry analysis and immunoblotting in Saos2 human osteogenic sarcoma cells. Curcumin and EF-24 inhibited the growth of Saos2 cells in a dose-dependent manner. The apparent potency of EF-24 was more than 3-fold higher that of curcumin. Treatment with curcumin or EF-24 resulted in nuclear condensation and fragmentation in the cells. The caspase-3/-7 activities were detected in living cells treated with curcumin or EF-24. Flow cytometry showed that the rate of apoptosis was increased by curcumin and EF-24 compared to the control. Curcumin and EF-24 promoted the proteolytic cleavages of procaspase-3/-7/-8/-9 with increases in the amount of cleaved caspase-3/-7/-8/-9. The curcumin- or EF-24-induced apoptosis in the Saos2 cells was mediated by the expression of Fas and activation of caspase-8, caspase-3 and poly(ADP-ribose) polymerase. Immunoblotting revealed the Bid and Bcl-2 proteins to be downregulated, and truncated-Bid, Bax and p53 proteins to be upregulated by curcumin and EF-24. Curcumin and EF-24 increased the Bax/Bcl-2 ratio significantly. These results suggest that the curcumin and EF-24 inhibit cell proliferation and induce apoptotic cell death in Saos2 human osteogenic sarcoma cells via both the mitochondria-mediated intrinsic pathway and the death receptor-mediated extrinsic pathway, and may have potential properties for anti-osteosarcoma drug discovery.

• Authors: Seok-Jin Yang, Seul Ah Lee, Min-Gyeong Park, Jae-Sung Kim, Sun-Kyoung Yu, Chun Sung Kim, Jin-Soo Kim, Su-Gwan Kim, Ji-Su Oh, Heung-Joong Kim, Hong Sung Chun, Yong Hwan Kim, Do Kyung Kim
• Journal: Oncology reports (Oncol Rep) Vol. 31 Issue 5 Pg. 2286-92 (May 2014) ISSN: 1791-2431 [Electronic] Greece
• PMID: 24604218 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 3,5-bis(2-fluorobenzylidene)piperidin-4-one
• Antineoplastic Agents
• BAX protein, human
• BH3 Interacting Domain Death Agonist Protein
• BID protein, human
• Benzylidene Compounds
• Fas-Associated Death Domain Protein
• Piperidones
• Tumor Suppressor Protein p53
• bcl-2-Associated X Protein
• Poly(ADP-ribose) Polymerases
• Caspase 3
• Caspase 7
• Caspase 8
• Caspase 9
• Curcumin

Topics

• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• BH3 Interacting Domain Death Agonist Protein (biosynthesis)
• Benzylidene Compounds (pharmacology)
• Bone Neoplasms (drug therapy)
• Caspase 3 (biosynthesis, metabolism)
• Caspase 7 (biosynthesis, metabolism)
• Caspase 8 (metabolism)
• Caspase 9 (metabolism)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Curcumin (pharmacology)
• Enzyme Activation (drug effects)
• Fas-Associated Death Domain Protein (biosynthesis)
• Humans
• Osteosarcoma (drug therapy)
• Piperidones (pharmacology)
• Poly(ADP-ribose) Polymerases (biosynthesis)
• Tumor Suppressor Protein p53 (biosynthesis)
• bcl-2-Associated X Protein (biosynthesis)