The Korea Journal of Herbology (대한본초학회지)

The Korea Association of Herbology (대한본초학회)
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Effect of Dangguibohyultang and its combinations on apoptosis in human colorectal adenocarcinoma HCT116 cells

당귀보혈탕(當歸補血湯)의 배합비율에 따른 대장암 세포주 HCT116의 세포사멸 효과

  • Kim, Byung-Wan (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University) ;
  • Yun, Hyun-Joung (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University) ;
  • Jeon, Hyeon-Suk (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University) ;
  • Yun, Hyung-Joong (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University) ;
  • Kim, Chang-Hyun (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University) ;
  • Park, Sun-Dong (Department of Herbal pharmacology, Collage of Oriental Medicine, Dongguk University)
  • 김병완 (동국대학교 한의과대학 방제학교실) ;
  • 윤현정 (동국대학교 한의과대학 방제학교실) ;
  • 전현숙 (동국대학교 한의과대학 방제학교실) ;
  • 윤형중 (동국대학교 한의과대학 방제학교실) ;
  • 김창현 (동국대학교 한의과대학 방제학교실) ;
  • 박선동 (동국대학교 한의과대학 방제학교실)
  • Published : 2006.06.30
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Abstract

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

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