title: 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.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Seok-Jin
authors.initials: SJ
authors.lastname: Yang
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Seul Ah
authors.initials: SA
authors.lastname: Lee
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Min-Gyeong
authors.initials: MG
authors.lastname: Park
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Sun-Kyoung
authors.initials: SK
authors.lastname: Yu
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Chun Sung
authors.initials: CS
authors.lastname: Kim
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Su-Gwan
authors.initials: SG
authors.lastname: Kim
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Ji-Su
authors.initials: JS
authors.lastname: Oh
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Heung-Joong
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Hong Sung
authors.initials: HS
authors.lastname: Chun
authors.affiliation: Korea Institute of Planning and Evaluation for Technology in Food, Agriculture Forestry and Fisheries, Gyeonggi-do 431-810, Republic of Korea.
authors.forename: Yong Hwan
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Oral Biology Research Institute, Chosun University, Gwangju 501-75, Republic of Korea.
authors.forename: Do Kyung
authors.initials: DK
authors.lastname: Kim
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D051023:BH3 Interacting Domain Death Agonist Protein / Q000096:biosynthesis; D001597:Benzylidene Compounds / Q000494:pharmacology*; D001859:Bone Neoplasms / Q000188:drug therapy*; D053148:Caspase 3 / Q000096:biosynthesis / Q000378:metabolism; D053179:Caspase 7 / Q000096:biosynthesis / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003474:Curcumin / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D053200:Fas-Associated Death Domain Protein / Q000096:biosynthesis; D006801:Humans; D012516:Osteosarcoma / Q000188:drug therapy*; D010881:Piperidones / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis
chemical_list: C524042:3,5-bis(2-fluorobenzylidene)piperidin-4-one; D000970:Antineoplastic Agents; C494826:BAX protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D001597:Benzylidene Compounds; D053200:Fas-Associated Death Domain Protein; D010881:Piperidones; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D053179:Caspase 7; D053181:Caspase 8; D053453:Caspase 9; D003474:Curcumin