Journal of Chest Surgery

The Korean Society for Thoracic and Cardiovascular Surgery (대한심장혈관흉부외과학회)
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Effects of Tobacco-Specific Carcinogen on Protein Kinase C Isoforms

흡연특이성 발암물질이 특정 Protein Kinase C Isoform에 미치는 영향

  • Kang, Hyung-Seok (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu) ;
  • Ko, Moo-Sung (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu) ;
  • Park, Ki-Sung (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu) ;
  • Lee, Sub (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu) ;
  • Jheon, Sang-Hoon (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu) ;
  • Kwon, Oh-Choon (Dept. of Thoracic and Cardiovascular Surgery, School of Medicine, Catholic University of Daegu)
  • 강형석 (대구가톨릭대학교 의과대학 흉부외과학교실) ;
  • 고무성 (대구가톨릭대학교 의과대학 흉부외과학교실) ;
  • 박기성 (대구가톨릭대학교 의과대학 흉부외과학교실) ;
  • 이섭 (대구가톨릭대학교 의과대학 흉부외과학교실) ;
  • 전상훈 (대구가톨릭대학교 의과대학 흉부외과학교실) ;
  • 권오춘 (대구가톨릭대학교 의과대학 흉부외과학교실)
  • Published : 2003.09.01
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Abstract

Cigarette smoking is the leading cause of the lung cancer. However, mechanism of action underlying the carcinogenesis in the lung still remains to be elucidated. The present study attempted to look into the carcinogenic potential of tobacco-specific nitrosamine, NNK (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone) and the effects of protein kinase C (PKC) isoforms in an immortalized human epithelial cell model. Material and Method: Immortalized human epithelial cells were exposed with NNK and examined for its carcinogenic potential as measured by saturation density, soft-agar colony formation, and cell aggregation assay. The specific isoform of PKCs involved in the cellular transformation was analysed through western blot with monoclonal antibody and measured separately in cytosolic fraction and membrane fraction. Result: Human epithelial cells exposed with NNK showed prominent carcinogenic potential in saturation density, soft agar colony formation, and cell aggregation assay. PKC isoform analysis results are as follows: PKC- $\alpha$ showed significant translocation of protein levels from cytosolic fraction to membrane fraction, as analyzed by immunoblot. PKC- $\varepsilon$ showed a dose-dependent increase of translocation. PKC- λ was not affected by NNK treatment. Conclusion: The study demonstrated that there was a certain specificity in the patterns of isoform induction following chemical carcinogen exposure. Thus, it is suggested that identification of specific isoform be a clue to find target molecules in the carcinogenesis.

폐암의 주된 원인으로 알려진 흡연은 그 악성세포 발현기전이 아직 정확히 규명된 바 없다. 이에 저자들은 흡연에 의한 발암성의 지표로 흡연 중에 특이적으로 존재하는 강력한 발암물질인 NNK(4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone)를 이용하여 흡연에 따른 폐암의 발생과 그 Protein kinase C (PKC) isoform과 관련된 기전에 관한 연구를 시도하였다. 대상 및 방법: 인체 상피세포를 NNK에 노출시킨 후 saturation density, soft agar colony formation, cell aggregation 및 foci의 출현 등의 양상을 파악하여 세포 발암성 여부를 관찰하였으며 NNK를 15분간 노출시킨 후 PKC의 변화는 세포 내 PKC isoform의 양을 cytosolic fraction과 membrane fraction으로 분리하여 측정하여 분석하였다. 결과: NNK 투여군에서 saturation density, soft agar colony formation, cell aggregation 및 foci의 출현 시기 등의 세포 발암성을 뚜렷이 나타내었으며 PKC isoform분석의 경우 PKC-$\alpha$의 membrane fraction의 뚜렷한 증가를 보였으며 이러한 활성은 용량-의존적인 형태를 유지하였다. PKC-$\varepsilon$은 NNK 처리 시 용량-의존적으로 cytosol fraction의 감소 및 membrane fraction의 증가를 뚜렷하게 보였고 NNK에 의한 PKC-λ의 변화는 감지되지 않았다. 결론: 본 연구는 화학적 발암물질인 NNK가 인체발암화에 관여함을 재차 확인하면서 초기 과정에 관여하는 PKC isoform의 변화를 분석함으로써 total PKC활성이 아닌 isoform 각각에 대한 변화를 확인하였다는 점에서 앞으로 인체상피세포 기원의 폐암 생성 기전 연구에 기여할 것으로 생각한다.

Keywords

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