Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Induction of Apoptosis by Piceatannol in YD-15 Human Oral Cancer Cells

피세아타놀에 의한 YD-15 구강암세포의 세포자가사멸 유도 효과

  • Lee, Hae-Nim (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Jang, Hye-Yeon (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Hyeong-Jin (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Shin, Seong-Ah (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Choo, Gang-Sik (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Park, Byung-Kwon (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Byeong-Soo (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University)
  • 이해님 (공주대학교 특수동물학과) ;
  • 장혜연 (공주대학교 특수동물학과) ;
  • 김형진 (공주대학교 특수동물학과) ;
  • 신성아 (공주대학교 특수동물학과) ;
  • 추강식 (공주대학교 특수동물학과) ;
  • 박병권 (공주대학교 특수동물학과) ;
  • 김병수 (공주대학교 특수동물학과) ;
  • 정지윤 (공주대학교 특수동물학과)
  • Received : 2015.03.19
  • Accepted : 2015.04.21
  • Published : 2015.07.31
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Abstract

Piceatannol (trans-3,4,3',5'-trihydroxystilbene), a natural stilbene, is an analogue of resveratrol. In the present study, possible mechanisms by which piceatannol exerts its pro-apoptotic action in cultured human oral cancer YD-15 cells were investigated. To investigate whether or not piceatannol has effects on cancer cell viability, human oral YD-15 cells were treated with piceatannol (0, 50, and $100{\mu}M$). Piceatannol treatment ($100{\mu}M$) showed the strongest inhibition of cell proliferation and reduced cell viability in a dose-dependent manner. Chromatin condensation detected by DAPI staining significantly increased in a concentration-dependent manner, indicating apoptosis. Piceatannol treatment activated initiator Bax (pro-apoptotic) and cPARP in a concentration-dependent manner. Further, piceatannol induced down-regulation of Bcl-2 (anti-apoptotic). We also evaluated the activity of piceatannol against oral cavity cancer tumors in mice. Piceatannol-treated nude mice bearing YD-15 xenograft tumors exhibited significantly reduced tumor volume and weight due to the potent effect of piceatannol on tumor cell apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Immunohistochemistry staining showed elevated expression of cleaved-caspase-3 as well as reduced expression of Ki-67 in the piceatannol-treated group. Therefore, piceatannol can be developed as a cancer preventive medicine due to its growth inhibitory effects and induction of apoptosis in human oral cancer cells.

Resveratrol 유도체의 일종으로 stilbene 계열 물질인 piceatannol은 암세포의 증식을 억제하고 apoptosis를 유발하는 것으로 알려져 있다. 본 연구에서는 YD-15 인체 구강암세포를 대상으로 piceatannol에 의한 암세포 증식 억제와 연관된 부가적인 기전연구를 실시하였다. Piceatannol이 암세포 성장에 미치는 영향을 확인하기 위하여 구강암세포주 YD-15에 piceatannol을 0, 6.25, 12.5, 25, 50, 100, $200{\mu}M$의 농도로 처리하고 MTT assay를 수행한 결과 piceatannol은 농도 의존적으로 세포 성장을 억제하였다. Piceatannol에 의한 암세포 증식 억제 효과가 apoptosis에 의한 것인지 확인하기 위해 DAPI 염색을 수행한 결과 apoptotic body와 세포질 응축이 농도 의존적으로 증가하는 것을 확인하였다. Western blotting의 결과 piceatannol은 Bax와 cleaved-PARP 단백질의 발현을 농도 의존적으로 증가시키고 Bcl-2 단백질 발현을 감소시켰다. In vivo 실험에서는 누드마우스에 YD-15 구강암세포를 이종이식한 후 3주간의 piceatannol 복강투여를 통해 이식된 종양의 크기를 측정한 결과 piceatannol 처치군이 대조군에 비해 유의적으로 종양 부피가 감소하였고, 종양조직을 이용해 TUNEL assay를 수행한 결과 piceatannol 처치군에서 TUNEL-positive cell이 더 많이 관찰되었다. 또한 종양조직을 이용해 IHC stain을 수행하여 cleaved-caspase-3와 Ki-67을 확인한 결과 piceatannol 처치군에서 cleaved-caspase-3가 증가하는 경향을 보였고 Ki-67은 감소하는 경향을 보였다. 따라서 piceatannol은 인체 구강암세포에서 apoptosis를 유도시키고 종양 형성을 억제함으로써 구강암 치료제로의 개발 가능성이 있는 것으로 확인하였다.

Keywords

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