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 Citrus grandis Osbeck Peel (CGP) Extract in HL60 Cells

당유자 과피 추출물에 의한 HL60 세포의 Apoptosis 유도

  • Hyon, Jae-Seok (Dept. of Food and Nutrition, Jeju College of Technology) ;
  • Kang, Sung-Myung (School of Marine Biomedicinal Sciences, Jeju National University) ;
  • Kim, Areum-Daseul (School of Marine Biomedicinal Sciences, Jeju National University) ;
  • Oh, Myung-Cheol (Dept. of Tourism Hotel Culinary Art, Jeju College of Technology) ;
  • Oh, Chang-Kyung (Dept. of Tourism Hotel Culinary Art, Jeju College of Technology) ;
  • Kim, Dong-Woo (Central Research Center, Natural F&P Co., Ltd.) ;
  • Jeon, You-Jin (School of Marine Biomedicinal Sciences, Jeju National University) ;
  • Kim, Soo-Hyun (Dept. of Food Bioengineering, Jeju National University)
  • 현재석 (제주산업정보대학 식품영양과) ;
  • 강성명 (제주대학교 해양의생명과학부) ;
  • 김아름다슬 (제주대학교 해양의생명과학부) ;
  • 오명철 (제주산업정보대학 관광호텔조리과) ;
  • 오창경 (제주산업정보대학 관광호텔조리과) ;
  • 김동우 ((주)네추럴 F&P 중앙연구소) ;
  • 전유진 (제주대학교 해양의생명과학부) ;
  • 김수현 (제주대학교 식품생명공학과)
  • Published : 2009.10.31
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Abstract

In the present study, we investigated the anti-proliferation activity of Citrus grandis Osbeck peel (CGP) in HL60 (human promyelocytic leukemia) cells. It was found that 80% ethanol extract of CGP could inhibit the cell growth in a dose-dependent manner ($250{\sim}1,000{\mu}g/mL$), which was associated with morphological changes and apoptotic cell death such as depolarized mitochondrial membrane, formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. The results indicate that CGP extract inhibits the growth of HL60 cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl family proteins and increase the activation of caspase-3 and PARP. Therefore, it is suggested that CGP has the potential to provide a remarkable natural defense against the proliferation of HL60 cells.

당유자 과피(GGP) 80% 에탄올 추출물을 4종의 암세포에 (피부암, 대장암, 유방암 및 혈액암) 처리하여 증식 억제 활성을 측정한 결과, 혈액암 HL60 세포에서 높은 증식 억제 활성을 보였다. 이에 CGP 추출물이 HL60 세포에 대한 apoptosis 유도에 따른 세포 증식 억제 활성을 조사하였다. Apoptosis 유도의 첫 단계인 막 투과성을 측정한 결과, confocal image와 flow cytometry에서 CGP를 처리하였을 때 탈분극 현상에 따른 막 투과성이 증가하였고 세포내 핵을 hoechst 33342를 이용하여 염색하였을 때 apoptosis가 일어났을 때 나타나는 전형적인 형태의 apoptotic body가 농도 의존적으로 증가하는 것을 확인할 수 있었으며 flow cytometry를 통하여 세포 주기를 측정하였을 때 DNA-hypodiploid 형태의 sub-G1가 CGP 농도 의존적으로 증가하는 것을 확인할 수 있었다. Apoptosis 유도 기전을 western blot으로 측정한 결과를 보면, CGP 추출물을 혈액암 HL60 세포에 처리하였을 때 Bcl family의 anti-apoptotic Bcl-2 단백질의 감소와 pro-apoptotic Bax 단백질의 증가로 인하여 하위 기전인 caspase-3가 활성화되었으며, 이 활성화로 인하여 apoptosis 유도에 직접적으로 관여하는 PARP 단백질을 활성화시키면서 apoptosis를 유도하였다. 따라서 당유자 과피는 항암과 관련되어진 기능성식품 및 소재 개발 원료로서 개발이 가능하리라고 사료된다.

Keywords

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