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Apoptotic Effect of Co-Treatment with Valproic Acid and 17AAG on Human Osteosarcoma Cells

Valproic acid와 17AAG의 병용처리가 사람골육종세포에 미치는 세포자멸사 효과에 대한 연구

  • Park, Jun-Young (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Park, Se-Jin (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, In-Ryoung (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Jeong, Sung-Hee (Department of Oral Medicine, School of Dentistry, Pusan National University) ;
  • Ko, Myung-Yun (Department of Oral Medicine, School of Dentistry, Pusan National University) ;
  • Ahn, Yong-Woo (Department of Oral Medicine, School of Dentistry, Pusan National University)
  • 박준영 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 박세진 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 김인령 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 박봉수 (부산대학교 치의학전문대학원 구강해부학교실) ;
  • 정성희 (부산대학교 치의학전문대학원 구강내과학교실) ;
  • 고명연 (부산대학교 치의학전문대학원 구강내과학교실) ;
  • 안용우 (부산대학교 치의학전문대학원 구강내과학교실)
  • Received : 2010.12.04
  • Accepted : 2011.01.30
  • Published : 2011.03.30

Abstract

Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent. And it is known that antitmor activity of VPA is associated with its targeted at histone deacetylases. 17AAG, Inhibition of HSP90 leads to the proteasome degradation of the HSP90 client proteins, such as Akt, Raf/Ras, Erk, VEGF, cyclin D and p53, and causes potent antitumor activity. It is reported that 17AAG-induced HSP90 inhibition results in prevention of cell proliferation and induction of apoptosis in several types of cancer. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and the HSP90 inhibitor, 17AAG on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and 17AAG showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-3, caspase-7 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or 0.5 ${\mu}M$ 17AAG for 48 h did not induce apoptosis, the co-treatment with them induced prominently apoptosis. Therefore our data in this study provide the possibility that combination therapy with VPA and 17AAG could be considered as a novel therapeutic strategy for human osteosarcoma.

Acknowledgement

Supported by : Pusan National University

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