생명과학회지 (Journal of Life Science)

  • 제17권10호
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  • Pages.1447-1451
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  • 2007
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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종양억제유전자 p53 결손 인체간암세포에서 Pectenotoxin-2에 의한 Apoptosis 유도

Apoptotic Cell Death by Pectenotoxin-2 in p53-Deficient Human Hepatocellular Carcinoma Cells

  • 신동역 (동의대학교 한의과대학 생화학교실) ;
  • 김기영 (제주대학교 해양과학대학 해양과학부) ;
  • 최병태 (부산대학교 한의학전문대학원 해부학교실) ;
  • 강호성 (부산대학교 자연과학대학 분자생물학과) ;
  • 정지형 (부산대학교 약학대학 약학과) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Shin, Dong-Yeok (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Kim, Gi-Young (Faculty of Applied Marine Science, Cheju National Univeristy) ;
  • Choi, Byung-Tae (Department of Anatomy, School of Oriental Medicine, Pusan National University) ;
  • Kang, Ho-Sung (Department of Molecular Biology, Pusan National University) ;
  • Jung, Jee-H. (Department of Pharmacy, Pusan National University) ;
  • Choi, Yung-Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine)
  • 발행 : 2007.10.30
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초록

해양생물 유래 항암활성을 가지는 천연물의 탐색과정에서 해면동물에서 유래된 PTX-2는 p53 결손 암세포에서 세포독성 효과가 높은 것으로 보고된 바 있다. 본 연구에서는 인체 간암세포 모델을 이용하여 PTX-2의 효능을 조사한 결과는 p53 결손 Hep3B 세포에서 p53 정상 HepG2에 비하여 항암활성이 매우 높았으며, 이는 apoptosis 유발과 연관성이 있음을 확인하였다. PTX-2에 의한 Hep3B 세포의 apoptosis 유발은 DFF family의 발현 변화, pro-apoptotic Bax 및 Bcl-xS 단백질의 발현 증가, caspases (-3, -8 및 -9)의 활성화 등이 관여함을 알 수 있었다. PTX-2는 또한 Hep3B 세포에서 AKT 및 ERK1/2의 활성화를 유도하였으며, caspase-3, AKT 및 ERK1/2의 특이적 저해제에 의하여 PTX-2에 의한 세포증식 억제 효능이 유의적으로 감소되었다. 본 연구는 PTX-2에 의한 Hep3B 세포에서의 apoptosis 유도에 AKT 및 ERK1/2 신호 전달 경로가 중요한 역할을 하고 있음을 보여주는 결과이다.

Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

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