Radiation Oncology Journal

The Korean Society for Radiation Oncology (대한방사선종양학회)
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Regulatory Mechanism of Radiation-induced Cancer Cell Death by the Change of Cell Cycle

세포주기 변화에 타른 방사선 유도 암세포 사망의 조절기전

  • Jeong Soo-Jin (Department of Radiation Oncology Dong-A University Hospital) ;
  • Jeong Min-Ho (Department of Radiation Oncology Dong-A University Hospital) ;
  • Jang Ji-Yeon (Department of Radiation Oncology Dong-A University Hospital) ;
  • Jo Wol-Soon (Department of Radiation Oncology Dong-A University Hospital) ;
  • Nam Byung-Hyouk (Department of Radiation Oncology Dong-A University Hospital) ;
  • Jeong Min-Za (Department of Radiation Oncology Dong-A University Hospital) ;
  • Lim Young-Jin (Department of Radiation Oncology Dong-A University Hospital) ;
  • Jang Byung Gon (Department of Radiation Oncology Dong-A University Hospital) ;
  • Youn Seon-Min (Department of Radiation Oncology Dong-A University Hospital) ;
  • Lee Hyung Sik (Department of Radiation Oncology Dong-A University Hospital) ;
  • Hur Won Joo (Department of Radiation Oncology Dong-A University Hospital) ;
  • Yang Kwang Mo (Department of Radiation Oncology, Korea Institute of Radiological & Medical Sciences)
  • 정수진 (동아대학교 의과대학 방사선종양학과) ;
  • 정민호 (동아대학교 의과대학 방사선종양학과) ;
  • 장지연 (동아대학교 의과대학 방사선종양학과) ;
  • 조월순 (동아대학교 의과대학 방사선종양학과) ;
  • 남병혁 (동아대학교 의과대학 방사선종양학과) ;
  • 정민자 (동아대학교 의과대학 방사선종양학과) ;
  • 임영진 (동아대학교 의과대학 방사선종양학과) ;
  • 장병곤 (동아대학교 의과대학 방사선종양학과) ;
  • 윤선민 (동아대학교 의과대학 방사선종양학과) ;
  • 이헝식 (동아대학교 의과대학 방사선종양학과) ;
  • 허원주 (동아대학교 의과대학 방사선종양학과) ;
  • 양광모 (원자력의학원 방사선종양학과)
  • Published : 2003.12.01
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Abstract

Purpose : In our Previous study, we have shown the main cel1 death pattern Induced by irradiation or protein tyrosine kinase (PTK) inhibitors in K562 human myeiogenous leukemic cell line. Death of the cells treated with irradiation alone was characterized by mitotic catastrophe and typical radiation-induced apoptosis was accelerated by herblmycin A (HMA). Both types of cell death were inhibited by genistein. In this study, we investigated the effects of HMA and genistein on cell cycle regulation and its correlation with the alterations of radiation-induced cell death. Materials and Methods: K562 cells In exponential growth phase were used for this study. The cells were Irradiated with 10 Gy using 6 MeV Linac (200-300 cGy/min). Immediately after irradiation, cells were treated with 250 nM of HMA or 25 $\mu$N of genistein. The distributions of cell cycle, the expressions of cell cycle-related protein, the activities of cyclin-dependent kinase, and the yield of senescence and differentiation were analyzed. Results: X-irradiated cells were arrested In the G2 phase of the cell cycle but unlike the p53-positive cells, they were not able to sustain the cell cycle arrest. An accumulation of cells in G2 phase of first ceil-cycle post-treatment and an increase of cyclin Bl were correlated with spontaneous, premature, chromosome condensation and mitotic catastrophe. HMA induced rapid G2 checkpoint abrogation and concomitant p53-independent Gl accumulation. HMA-induced cell cycle modifications correlated with the increase of CDK2 kinase activity, the decrease of the expressions of cyclins I and A and of CDK2 kinase activity, and the enhancement of radiation-induced apoptosis. Genistein maintained cells that were arrested in the G2-phase, decreased the expressions of cyclin Bl and cdc25c and cdc25C kinase activity, increased the expression of pl6, and sustained senescence and megakaryocytic differentiation. Conclusion: The effects of HMA and genistein on the radiation-induced cell death of KS62 cells were closely related to the cell cycle regulatory activities. In this study, we present a unique and reproducible model in which for investigating the mechanisms of various, radiation-induced, cancer cell death patterns. Further evaluation by using this model will provide a potent target for a new strategy of radiotherapy.

목적: K562 세포의 방사선에 의한 세포 사망은 mitotic catastrophe 현상이 위주로 나타나지만 herbimycin A (HMA)에 의하여 apoptosis 반응이 촉진되는 반면 genisteln에 의하여 두 가지 형태의 세포사망이 모두 억제된다. 본 연구에서는 HMA와 genistein에 의한 K562세포의 방사선 유도 세포주기 조절 변화와 세포 사망 양상의 연관성을 조사하였다. 대상 및 방법: 지수증식기의 KS62 세포에 6 MV 선형가속기(Clinac 1,m C, Varian)를 이용하여 200~300 cGy/min의 선량률로 10 Gy를 균일하게 조사하였다. HMA와 genistein은 각각 250 nM와 25$\mu$M농도로 방사선 조사 후 즉시 투여하였다. 실험에서는 세포주기, 오절인자의 발현 및 활성, 노화 및 분화정도 등에 있어서의 시간에 따른 변화를 조사하였다. 결과: 방사선 단독조사에서 KS62세포는 G2기의 정체를 보였으나 정상적인 053을 가지는 세포와는 달리 지속적인 세포주기의 정체를 보이지 않았다. G2정체가 유지되는 동안 cyclin Bl의 점진적인 증가를 관찰할 수 있었으며, 이는 염색체의 복제가 완료되지 않은 상태에서 M기로 진행하여 미성숙한 염색체 응축과 mitotic catastrophe 현상이 나타나는 것과 일치한다. 방사선 조사와 함께 HMA를 투여한 경우에는 G2정체가 빠르게 해소되었으며 동시에 Gl기에서 세포가 정체되는 양상을 보였다. 세포주기 조절인자 cdc2 kinase 활성 증가와 cyclln I와 A 발현 및 CDK2 활성의 감소 등의 현상으로 설명되며, 이는 apoptosis의 증가와 연관성을 갖는다. 반면 genistein의 경우에는 cyclin Bl과 떨cfsc 발현 및 cdc2활성이 모두 감소하는 등 G2정체를 계속 유지하였다. 이와 함께 방사선에 의한 노화와 megakaryocyte로의 분화도 지속되는 것을 관찰할 수 있었다. 결론: HMA와 genistein에 의한 KS62세포의 방사선 유도 세포사망의 변화는 세포주기 조절과 밀접하게 연관되어 있음을 확인하였다. 이는 다양한 방사선 유도 세포사망의 기전을 이해하는 데 독창적인 모델을 제공하며, 방사선을 이용한 암 치료법의 개발에 새로운 표적을 제공할 수 있을 것이다.

Keywords

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