Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Evaluation of Cell Death and the Reduction of ERK Phosphorylation in Non-Small Cell Lung Cancer Cells after Exposure to Sodium Butyrate

Sodium butyrate 노출에 의한 비소세포폐암 세포의 세포사멸과 extracellular signal-regulated kinase 인산화의 감소

  • Park, Ji-Eun (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Lee, Seung-Gee (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Lim, Hyun-Ju (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Kim, Ji-Young (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Chung, Jin-Yong (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Kim, Yoon-Jae (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Lee, Chang-Hun (Department of Pathology, Pusan National University School of Medicine) ;
  • Lee, Min-Ki (Department of Internal Medicine, Pusan National University School of Medicine) ;
  • Yoo, Ki-Soo (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Yoo, Young-Hyun (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University) ;
  • Kim, Jong-Min (Department of Anatomy and Cell Biology, College of Medicine, Dong-A University)
  • 박지은 (동아대학교 의과대학 해부학교실) ;
  • 이승기 (동아대학교 의과대학 해부학교실) ;
  • 임현주 (동아대학교 의과대학 해부학교실) ;
  • 김지영 (동아대학교 의과대학 해부학교실) ;
  • 정진용 (동아대학교 의과대학 해부학교실) ;
  • 김윤재 (동아대학교 의과대학 해부학교실) ;
  • 이창훈 (부산대학교 의과대학 병리학교실) ;
  • 이민기 (부산대학교 의과대학 내과학교실) ;
  • 유기수 (동아대학교 의과대학 해부학교실) ;
  • 유영현 (동아대학교 의과대학 해부학교실) ;
  • 김종민 (동아대학교 의과대학 해부학교실)
  • Published : 2009.09.30
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Abstract

Histone deacetylase inhibitor (HDACI) is a new promising candidate as an antineoplastic agent for the treatment of solid and hematologic malignancies. In order to evaluate cell death and to elucidate the related mechanism(s) in NSCLC cells after HDACI, sodium butyrate (SB), a representative HDACI, was used to treat H460 cells for 48 hrs. SB exposure resulted in a significant reduction of cell viability at concentrations below 7.5 mM, and about 50% of cell death occurred at 20 mM. The types of cell death induced by SB were both apoptosis and necrosis, evaluated by Annexin-V staining combined with propidium iodide. SB treatment significantly evoked G2/M cell cycle arrest and subsequently induced cell death with caspase-dependent manner. While ERK protein content was not altered after SB, phosphorylated forms of ERK were markedly reduced. Taken together, SB is significantly able to induce cell death in NSCLC cell line H460, and it is suggested that the reduction of ERK phosphorylation might be closely involved in the cancer cell death mechanism initiated by HDACI.

히스톤 디아세틸라이제 저해제(HDACI)는 최근에 새로운 미래형 항암제로 주목을 받고 있으며 다른 항암요법 및 치료제와의 병용치료에도 효과적으로 적용될 수 있을 것으로 기대하고 있다. HDACI은 다양한 조직기원의 암세포에서 증식억제 및 세포사멸 유도능이 시험되어 왔으나 비소세포폐암 세포에서 그 작용 및 기전이 명확히 조사된 바가 없다. 본 연구는 HDACI 중의 하나인 sodium butyrate (SB)를 비소세포폐암 세포주인 H460에 처리하여 세포생존율, 세포주기 분석, 세포사멸도를 평가하고, 이와 관련하여 세포사멸 관련 단백질, p53, ERK의 변화를 조사하고자 하였다. 3가지 다른 농도(2.5, 7.5, 20 mM)의 SB에 H460 세포가 48시간 노출되었을 때, 세포 생존율은 농도의존적으로 감소되었으나 7.5 mM 이상의 농도에서 대조군에 비해 유의한 생존을 감소를 보였고, 20 mM에서 생존을 50% 전후를 나타냈다. SB노출은 H460 세포의 사멸을 유발하였는데, 세포사의 유형은 아포토시스와 괴사가 동시에 발생함이 Annexin-V 분석으로 확인되었다. H460 세포에서 SB에 의해 유발되는 뚜렷한 세포주기의 변화양상은 G2/M기정지였으며, 이러한 세포주기의 지연현상으로 세포사멸이 초래되는 것으로 생각된다. SB처리는 아포토시스 발생관련 효소인 caspase-3과 caspase-7의 활성화를 유발하였으며, 이에 의한 PARP 단백 질의 분절화도 관찰되었다. 동시에, 항세포사별 단백질인 XIAP의 단백질 함량은 감소함을 보였다. SB 노출에 의한 세포주기의 G2/M기의 정지현상과 관련하여는 p53 단백질의 증가가 주목할 만한 하였다. SB의 H460세포에의 처리는 일반 ERK단백질의 함량 변화를 유도하지 않았으나, 인산화형의 ERK는 SB처리농도에 의존적으로 그 단백질 함량이 감소하였다. 이는 ERK가 비소세포폐암 세포인 H460에서 세포생존 및 유지와 관련된 단백질의 인산화에 계속적으로 관여하고 있다는 사실을 암시한다. 즉, SB의 처리는 ERK의 인산화를 유의하게 억제하는 기전과 관련이 있을 가능성이 높다고 추측된다. 향후 SB의 노출에 의한 PERK 감소 기전에 대한 연구가 추가적으로 진행되면 SB의 더욱 효율적인 암세포 사멸 유도 전략수립에 도움을 줄 수 있을 것이라 예상된다.

Keywords

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