Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Induction of Apoptosis by Citri Pericarpium Methanol Extract through Reactive Oxygen Species Generation in U937 Human Leukemia Cells

진피 메탄올 추출물의 활성산소종 생성을 통한 인체 백혈병 세포의 apoptosis 유발

  • Kim, Ga Hee (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Lee, Moon Hee (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Han, Min Ho (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Park, Cheol (Department of Molecular Biology, College of Natural Sciences, Dongeui University) ;
  • Hong, Su Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Choi, Yung Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine)
  • 김가희 (동의대학교 한의과대학 생화학교실) ;
  • 이문희 (동의대학교 한의과대학 생화학교실) ;
  • 한민호 (동의대학교 한의과대학 생화학교실) ;
  • 박철 (동의대학교 자연과학대학 분자생물학과) ;
  • 홍수현 (동의대학교 한의과대학 생화학교실) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Received : 2013.08.22
  • Accepted : 2013.08.28
  • Published : 2013.08.30
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Abstract

Citri Pericarpium is one of the most commonly used traditional herbal medicines in Korea, China, and Japan. Its extracts have many properties including the treatment of indigestion and inflammatory respiratory syndromes such as bronchitis and asthma. However, the underlying molecular mechanisms of anti-cancer activity and molecular targets are not fully understood. In this work, we investigated the anti-proliferative activity of Citri Pericapium (EMCP) methanol extract on reactive oxygen species (ROS) production and the association of these effects with apoptotic cell death using U937 human leukemia cells in vitro. EMCP treatment decreased cell proliferation in a dose-dependent manner following an increase of the sub-G1 phase, the down-regulation of Bax proteins, the activation of caspases, the degradation of poly (ADP-ribose) polymerase proteins (PARP), and the induction of ROS generation. However, the quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the EMCP-induced apoptosis effects. In addition, heme oxygenase-1 expression also recovered by inhibiting the nuclear translocation of phosphorylated NF-E2-related factor 2. Taken together, our data indicate that ROS are involved as key mediators in the early molecular events in the EMCP-induced apoptotic pathway.

진피(Citri Pericarpium)의 항암작용 기전 해석을 위하여 U937 백혈병 세포의 apoptosis 유발에 미치는 메탄올 추출물(EMCP)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화, ROS의 생성 변화를 조사하였다. EMCP 처리에 의한 U937 세포의 증식 억제는 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 Flow cytometry 분석에 의한 Sub-G1기 세포 빈도의 증가로 확인하였다. EMCP 처리에 의한 apoptosis 유도에서 Bax 발현 증가, caspases의 활성 및 PARP의 단편화 등이 동반되었으며, ROS 생성의 증가와 연관성이 있었다. 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포 내 항산화 효소인 HO-1의 발현이 EMCP의 처리에 의해 증가되었으나 ROS 생성 억제제인 NAC의 전처리에 의해 감소된 HO-1의 발현은 전사인자인 Nrf2의 핵으로의 이동 억제와 관련되어 있었다. 이상의 결과에서 EMCP 처리에 의한 U937 세포의 apoptosis 유발에는 ROS 생성의 증가와 pNrf2에 의해 조절되는 HO-1의 발현 증가가 중요한 기전으로 작용한다는 것을 알 수 있었다. 이러한 자료는 진피의 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다.

Keywords

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