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 and Inhibition of Growth in Human Gastric Cancer by Piperine

Piperine에 의한 위암세포 AGS 증식 억제와 Apoptosis 유도

  • Shin, Seong-Ah (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Lee, Hae-Nim (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Choo, Gang-Sik (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, So-Jung (Department of Program in Cell Biology and Genetics, College of Medicine, Chungbuk National University) ;
  • Kim, Hyeong-Jin (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Park, Young-Seok (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) ;
  • Kim, Sang-Ki (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Lee, Hu-Jang (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University)
  • 신성아 (공주대학교 특수동물학과) ;
  • 이해님 (공주대학교 특수동물학과) ;
  • 추강식 (공주대학교 특수동물학과) ;
  • 김소정 (충북대학교 의과대학 의학과 생화학교실) ;
  • 김형진 (공주대학교 특수동물학과) ;
  • 박영석 (공주대학교 특수동물학과) ;
  • 박병권 (공주대학교 특수동물학과) ;
  • 김병수 (공주대학교 특수동물학과) ;
  • 김상기 (공주대학교 특수동물학과) ;
  • 이후장 (경상대학교 수의과대학 동물의학연구소) ;
  • 정지윤 (공주대학교 특수동물학과)
  • Received : 2016.07.18
  • Accepted : 2016.08.24
  • Published : 2016.11.30
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Abstract

Piperine [(E,E)-5-(3,4-methtylenedioxyphenyl)-2,4-pentadienolypiperidide] is a principal of Piperaceae, including Piper nigrum L. and Piper longum Linn., which has been used as a spice and traditional medicine. In this study, we investigated whether or not piperine has anti-cancer effects on AGS human gastric cancer cells. The results demonstrated that piperine not only inhibited proliferation using MTT assay but also induced apoptotic bodies using DAPI assay in a dose-dependent manner in response to piperine. Expression levels of p53, Bax (pro-apoptotic), cleaved caspase-9, and cleaved-PARP increased, whereas expression levels of Bcl-2, XIAP (anti-apoptotic), and Akt decreased in a dose-dependent manner compared with the control by western blotting analysis. To identify the connection between phospo-Akt and Bcl-2 family in response to piperine, LY249002 (Akt inhibitor) was treated with piperine ($150{\mu}M$). The results were shown that expression of phospo-Akt was reduced whereas expression of Bax and cleaved-PARP increased in a dose-dependent manner. These results indicate that piperine induced apoptosis in AGS cells and may serve as a chemopreventive or therapeutic agent for human gastric cancer.

본 연구는 후추과 식물의 주성분인 piperine이 위암세포 AGS의 항암 효과에 미치는 영향을 확인하기 위해 수행되었다. Piperine에 의한 위암세포 AGS의 생존율을 확인하기 위해 MTT assay를 수행한 결과 농도 의존적으로 암세포의 생존율이 감소하는 것을 확인하였다. 이러한 암세포의 생존율 억제가 apoptosis에 의한 효과인지를 확인하기 위해 DAPI staining을 실시한 결과 piperine을 처치한 군에서 apoptotic body와 염색질 응축이 증가하는 것을 확인하였다. MTT assay와 DAPI staining의 결과를 바탕으로 piperine이 위암세포 AGS에서 apoptosis와 관련한 단백질 발현 양상에 미치는 영향을 확인하기 위해 western blotting을 실시하였다. 그 결과 piperine에 의해 AGS 세포에서 apoptosis를 유도하는 p53, Bax의 발현은 농도 의존적으로 증가하였고, apoptosis를 억제하는 Bcl-2, XIAP의 발현은 농도 의존적으로 감소하였다. Apoptosis의 궁극적 단계인 caspase-9와 손상된 DNA를 복구하는 PARP의 분절은 증가하였으며, apoptosis를 방해하는 인자인 Akt의 인산화는 농도 의존적으로 감소하였다. Akt 억제제인 LY294002와 piperine을 병행 또는 단독으로 처치하여 western blotting을 진행한 결과 Bcl-2의 발현은 piperine을 단독으로 처치했을 때와 유사한 감소 경향을 보였지만, p-Akt의 발현은 대조군과 비교해 LY294002와 piperine을 단독으로 처치했을 때보다 병행했을 때 더욱 감소하였다. 이와 반대로 Bax와 cleaved-PARP의 발현은 강하게 증가하였다. 본 연구의 결과를 종합해볼 때 piperine이 위암세포 AGS에서 Akt 경로를 통해 apoptosis를 유도하는 것으로 여겨지며, 향후 위암 예방제나 치료제로의 개발 가능성이 있을 것으로 생각한다.

Keywords

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