Radiation Oncology Journal

  • 제21권3호
  • /
  • Pages.227-237
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  • 2003
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  • 2234-1900(pISSN)
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  • 2234-3156(eISSN)
대한방사선종양학회 (The Korean Society for Radiation Oncology)
권호 표지

K562 세포의 방사선 감수성 변화에 영향을 미치는 신호전달인자

Signal Transduction Factors on the Modulation of Radiosusceptibility in K562 Cells

  • 양광모 (원자력의학원 방사선종양학과) ;
  • 윤선민 (을지의과대학교 방사선종양학과) ;
  • 정수진 (동아대학교 의과대학 의과학 연구소) ;
  • 장지연 (동아대학교 의과대학 의과학 연구소) ;
  • 조월순 (동아대학교 의과대학 의과학 연구소) ;
  • 도창호 (동아대학교 의과대학 의과학 연구소) ;
  • 유여진 (동아대학교 의과대학 의과학 연구소) ;
  • 신영철 (동아대학교 의과대학 의과학 연구소) ;
  • 이형식 (동아대학교 의과대학 치료방사선과학교실) ;
  • 허원주 (동아대학교 의과대학 치료방사선과학교실) ;
  • 임영진 (동아대학교 의과대학 의과학 연구소) ;
  • 정민호 (동아대학교 의과대학 의과학 연구소)
  • Yang Kwang Mo (Korea Institute of Radiological & Medical Sciences) ;
  • Youn Seon-Min (Department of Radiation Oncology, Eulji University School of Medicine) ;
  • Jeong Soo-Jin (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Jang Ji-Yeon (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Jo Wol-Soom (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Do Chang-Ho (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Yoo Y대-Jin (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Shin Young-Cheol (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Lee Hyung Sik (Department of Radiation Oncology, Dong-A University Hospital, College of Medicine) ;
  • Hur Won Joo (Department of Radiation Oncology, Dong-A University Hospital, College of Medicine) ;
  • Lim Young-Jin (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine) ;
  • Jeong Min-Ho (The Institute of Medical Science (BK21 program), Dong-A University Hospital, College of Medicine)
  • 발행 : 2003.09.01
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초록

목적: 만성 골수성 백혈병 세포인 K562 세포주는 방사선 및 다양한 항암제에 대한 apoptosis에 저항성을 가진다. 지난 연구에서 K562 세포는 방사선에 대하여 내성반응을 보이며, 세포내 PTK의 작용을 억제하고자 방사선 조사와 함께 투여한 herbimycin A (HMA)에 의하여 방사선에 대한 apoptosis와 같은 감수성반응이 유도되는 반면, genistein에 의하여 방사선에 대한 apoptosis 반응이 저해됨을 확인하였다. 본 연구에서는 타이로신 인산화효소 억제에 의한 K562 세포의 방사선 반응변화를 조절하는 신호전달경로를 조사하였다. 대상 및 방법: K562 세포를 지수증식기의 세포들만 선택하여 실험에 이용하였다. 방사선조사는 6 MeV 선형가속기(Clinac 1800C, Varian)를 이 용하여 $200\~300$ cGy/min 선량률로 $0.5\~12 $ Gy를 균일하게 조사하였다. HMA와 genistein은 각각 $0.25/muM,\;25\muM$을 방사선 조사 후 즉시 투여하였다. 실험에서 신호전달 경로로 abl kinase, MAPK family, NF-kB, c-fos, c-myc, thymidine kinase1 (TK1) 등에서의 단백질 또는 유전자 발현 및 활성을 조사하였다. 또한 약제 투여에 따른 유전자 발현차이(differential gene expression)를 조사하였다. 결과: Abl kinase의 발현 및 활성 변화를 조사하였으나 PTK 저해제에 의한 방사선 유도 세포사의 변화와의 연관성을 찾을 수 없었다. 세포 생존 및 사멸의 신호전달체계에서 주요 조절과정인 MAPK family의 관여 여부 확인에서 방사선으로 인한 SAPK/JNK의 활성화의 유도가 관찰되었으나, PTK 저해제에 따른 변화는 없었으며, 또한 MAPK/ERK와 p38 MAPK 활성은 모든 조건에서 변함 없이 일정하였다. 전사인자 활성화에 대한 조사에서 방사선 조사와 함께 genistein을 투여한 경우에 NF-kB활성이 증가하였다. 유전자 발현 차이의 조사에서 genistein 투여에 의한 TK 1 유전자 발현 및 단백질 활성이 증가하였다. 결론: PTK 억제에 의한 K562 세포의 방사선에 대한 반응 변화는 bcrabl kinase 활성과는 무관하게 진행되며, MAPK family 경로 외의 다른 경로를 통한 전사인자 활성화 과정이 연관되어 있음을 확인하였다.

Purpose: The human chronic myelogenous leukemia cell line, K562, expresses the chimeric bcr-abl oncoprotein, whose deregulated protein tyrosine kinase activity antagonizes via DNA damaging agents. Previous experiments have shown that nanomolar concentrations of herbimycin A (HWA) coupled with X-irradiation have a synergistic effect in inducing apoptosis in the Ph-positive K562 leukemia cell line, but genistein, a PTK inhibitor, is non selective for the radiation-induced apoptosils on $p210^{bcr/abl}$ protected K562 cells. In these experiments, the cytoplasmic signal transduction pathways, the Induction on a number of transcription factors and the differential gene expression in this model were investigated. Materials and Methids: K562 cells in the exponential growth phase were used in this study. The cells were irradiated with 0.5-12 Gy, using a 6 Mev Linac (Clinac 1800, Varian, USA). Immediately after irradiation, the cells were treated with $0.25/muM$ of HMA and $25/muM$ of genistein, and the expressions and the activities of abl kinase, MAPK family, NF- kB, c-fos, c-myc, and thymidine kinase1 (TK1) were examined. The differential gene expressions induced by PTK inhibitors were also investigated. Results: The modulating effects of herbimycin A and genistein on the radiosensitivity of K562 cells were not related to the bcr-abl kinase activity. The signaling responses through the MAPK family of proteins, were not involved either in association with the radiation-induced apoptosis, which is accelerated by HMA, the expression of c-myc was increased. The combined treatment of genistein, with irradiation, enhanced NF- kB activity and the TK1 expression and activity. Conclusion: The effects of HMA and genistein on the radiosensitivity on the K562 cells were not related to the bcr-abl kinase activity in this study, another signaling pathway, besides the WAPK family responses to radiation to K562 cells, was found. Further evaluation using this model will provide valuable information for the optional radiosensitization or radioprotection.

키워드

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