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Induction of Apoptosis in Human Colon Carcinoma HCT116 Cells Using a Water Extract of Lepidium virginicum L.

콩다닥냉이 추출물에 의한 HCT116 대장암세포의 사멸 유도에 관한 연구

  • Chae, Yang-Hui (Dept. of Biology, Kyungsung University) ;
  • Shin, Dong-Yeok (Dept. of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Park, Cheol (Dept. of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Lee, Yong-Tae (Dept. of Pathology, Research Institute of Oriental Medicine, Dongeui University College of Oriental Medicine) ;
  • Moon, Sung-Gi (Dept. of Biology, Kyungsung University) ;
  • Choi, Yung-Hyun (Dept. of Biochemistry, Dongeui University College of Oriental Medicine)
  • 채양희 (경성대학교 자연과학대학 생물학과) ;
  • 신동역 (동의대학교 한의과대학 생화학교실) ;
  • 박철 (동의대학교 한의과대학 생화학교실) ;
  • 이용태 (동의대학교 한의과대학 생리학교실 및 한의학연구소) ;
  • 문성기 (경성대학교 자연과학대학 생물학과) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Received : 2011.03.11
  • Accepted : 2011.04.13
  • Published : 2011.05.31

Abstract

To examine the anti-cancer effects of Lepidium virginicum L., the anti-proliferative and pro-apoptotic effects of a water extract of L. virginicum leaves (WELVL) and of L. virginicum roots (WELVR) were investigated in HCT116 human colon carcinoma cells. The treatment of HCT116 cells with WELVL and WELVR resulted in the inhibition of growth and morphological changes in a concentration-dependent manner by inducing apoptosis. The growth inhibition and apoptosis induction by WELVR was stronger than that of WELVL thus, we determined that WELVR was the more optimal extract for this study. The increased apoptotic events in HCT116 cells caused by WELVR were associated with an up-regulation of Fas ligand, Bax, and Bad expression, a down-regulation of Bcl-2, Bcl-$_XL$, and Bid expression, and a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\psi}m$). WELVR treatment induced the proteolytic activation of caspase-3, -8, and -9, and the degradation of caspase-3 substrate proteins, such as poly (ADP-ribose) polymerase (PARP), ${\beta}$-catenin, and phospholipase C-${\gamma}1$ (PLC-${\gamma}1$). In addition, apoptotic cell death induced by WELVR was correlated with a down-regulation of inhibitors of the apoptosis protein (IAP) family, such as the X-linked inhibitor of apoptosis protein (XIAP), cIAP-1, and cIAP-2. These findings suggest that the WELVR-induced inhibition of cell proliferation is associated with the induction of apoptotic cell death. WELVR may be a potential chemotherapeutic agent for the control of HCT116 human colon carcinoma cells.

연구에서는 콩다닥냉이 추출물의 항암활성을 조사하기 위하여 잎 및 뿌리의 열수 추출물(WELVL 및 WELVR)이 HCT116 대장암세포의 증식 억제와 연관된 apoptosis 유도 기전에 관한 연구를 시도하였다. 본 연구의 결과에 의하면 HCT116 세포에 WELVL 및 WELVR을 처리하였을 경우에 유발되는 증식 억제 및 형태 변화는 apoptosis 유발과 밀접한 연관이 있었으며, 증식억제 및 apoptosis 유도 효과는 WELVL에 비하여 WELVR에서 높게 나타났다. 특히 WELVR에 의한 apoptosis 유발에는 FasL의 발현 증가를 통한 caspase-8의 활성화와 이로 인한 Bid 단백질의 단편화와 함께 Bcl-2 family의 발현 변화를 통한 mitochondria의 기능 이상과 이로 인한 caspase-9 및 -3의 활성화, 그리고 기질단백질들의 분해가 중요한 역할을 하는 것으로 나타났다. 또한 IAP family의 발현 감소로 인한 caspase의 활성 증가도 어느 정도 관여하는 것으로 생각되어진다. 따라서 WELVR 처리에 의하여 유발되는 apoptosis는 extrinsic pathway 및 intrinsic pathway를 모두 경유하는 multiple apoptotic pathway에 의하여 조절되는 것으로 생각되며, 이러한 결과들은 인체 암세포에서 콩다닥냉이의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 콩다닥냉이 추출물을 포함한 그와 유사한 항암제 후보물질들의 연구 기초자료로서 사용될 수 있을 것으로 생각된다.

Keywords

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