Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Gene Expression Profile of Lung Cancer Cells Following Photodynamic Therapy

폐암 세포주에서 광역학 치료에 의한 유전자 발현 분석

  • Sung, Ji Hyun (Department of Molecular and Cellular Biochemistry, College of Medicine, Kangwon National University) ;
  • Lee, Mi-Eun (Clinical Research Institute of Kangwon National University Hospital) ;
  • Han, Seon-Sook (Department of Internal Medicine, College of Medicine, Kangwon National University) ;
  • Lee, Seung-Joon (Department of Internal Medicine, College of Medicine, Kangwon National University) ;
  • Ha, Kwon-Soo (Department of Molecular and Cellular Biochemistry, College of Medicine, Kangwon National University) ;
  • Kim, Woo Jin (Department of Internal Medicine, College of Medicine, Kangwon National University)
  • 성지현 (강원대학교 의과대학 생화학교실) ;
  • 이미은 (강원대학교병원 임상의학연구소) ;
  • 한선숙 (강원대학교 의과대학 내과학교실) ;
  • 이승준 (강원대학교 의과대학 내과학교실) ;
  • 하권수 (강원대학교 의과대학 생화학교실) ;
  • 김우진 (강원대학교 의과대학 내과학교실)
  • Received : 2007.04.05
  • Accepted : 2007.06.07
  • Published : 2007.07.30
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Abstract

Background: Photodynamic therapy is a viable option for lung cancer treatment, and many studies have shown that it is capable of inducing cell death in lung cancer cells. However, the precise mechanism of this cell death has not been fully elucidated. To investigate the early changes in cancer cell transcription, we treated A549 cells with the photosensitizer DH-I-180-3 and then we illuminated the cells. Methods: We investigated the gene expression profiles of the the A549 lung cancer cell line, using a DEG kit, following photodynamic therapy and we evaluated the cell viability by performing flow cytometry. We identified the genes that were significantly changed following photodynamic therapy by performing DNA sequencing. Results: The FACS data showed that the cell death of the lung cancer cells was mainly caused by necrosis. We found nine genes that were significantly changed and we identified eight of these genes. We evaluated the expression of two genes, 3-phosphoglycerate dehydrogenase and ribosomal protein S29. The expressed level of carbonic anhydrase XII, clusterin, MRP3s1 protein, complement 3, membrane cofactor protein and integrin beta 1 were decreased. Conclusion: Many of the gene products are membrane-associated proteins. The main mechanism of photodynamic therapy with using the photosensitizing agent DH-I-180-3 appears to be necrosis and this may be associated with the altered production of membrane proteins.

연구배경: 광역학 치료는 폐암 치료에 실질적으로 이용 가능하며, 많은 연구들에서 폐암 세포에서 세포사멸을 일으킨다는 것이 이미 알려져 있다. 그러나 이 세포사멸의 기전은 아직 정확히 알려져 있지 않으며, 이에 암세포의 전사에서 초기 변화가 어떻게 일어나는 지를 알아보기 위하여 실험을 수행하였다. 방 법: 광과민성 물질인 DH-I-180-3으로 A549 세포에 처리를 하고 광역학 치료를 한 후 관찰하였다. 광역학 치료 후 DEG kit를 이용하여 폐암 세포주에서의 유전자 발현을 보았으며, 유세포 분석기를 이용하여 세포 사멸을 측정하였다. 광역학 치료 후 의미있는 변화를 보인 유전자는 염기서열분석으로 확인하였다. 결 과: 유세포분석 결과 폐암세포주는 대부분 세포괴사에 의하여 사멸되었다.광역학 치료 후, 9개의 유전자에서 명확한 변화가 있음을 발견했으며 이 중8개의 유전자를 밝혀내었다. 3-phosphoglycerate dehydrogenase와 리보솜 단백질 S29의 유전자 발현이 증가되어 있었으며, carbonic anhydrase XII, clusterin, MRP3s1 protein, complement 3, membrane cofactor protein, ${\beta}$-1 integrin의 유전자 발현은 감소되어 있었다. 결 론: 본 연구는 광과민성 물질인 DH-I-180-3을 이용한 광역학 치료에서 폐암 세포의 세포사멸의 주된 기전이 세포괴사에 의해 이루어 진 것임을 밝혀냈으며, 이와 관련된 유전자들 대부분이 막단백의 변화를 통해 이루어짐을 알 수 있었다.

Keywords

References

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