International Journal of Oral Biology

  • 제35권2호
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  • Pages.51-60
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  • 2010
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  • 1226-7155(pISSN)
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  • 2287-6618(eISSN)
대한구강생물학회 (The Korean Academy of Oral Biology)
권호 표지

Mechanism Underlying NaF-Induced Apoptosis in Human Oral Squamous Cell Carcinoma

  • Hur, Young-Joo (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, Do-Kyun (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Lee, Seung-Eun (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, In-Ryoung (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Jeong, Na-Young (Department of Anatomy, College of Medicine, Dong-A University) ;
  • Kim, Ji-Young (Department of Anatomy, College of Medicine, Dong-A University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, School of Dentistry, Pusan National University)
  • 투고 : 2010.05.06
  • 심사 : 2010.06.04
  • 발행 : 2010.06.30
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초록

Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor $p27^{KIP1}$. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.

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