Journal of the Korean Association of Oral and Maxillofacial Surgeons

The Korean Association of Oral and Maxillofacial Surgeons (대한구강악안면외과학회)
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PHOTOTOXIC EFFECTS OF LOW LEVEL LASER IRRADIATION ON HUMAN OSTEOSARCOMA CELLS

골육종세포에 미치는 레이저 조사의 광독성 효과

  • Son, Jang-Ho (Dept. of Oral & Maxillofacial Surgery, Medical College of Ulsan University) ;
  • Cho, Young-Chul (Dept. of Oral & Maxillofacial Surgery, Ulsan Dong Kang Hospital) ;
  • Ryu, Sung-Ho (Dept. of Oral & Maxillofacial Surgery, Medical College of Ulsan University) ;
  • Kim, Gyoo-Cheon (Dept. of Oral Anatomy, College of Dentistry, Pusan National University) ;
  • Sung, Iel-Yong (Dept. of Oral & Maxillofacial Surgery, Medical College of Ulsan University) ;
  • Park, Bong-Soo (Dept. of Oral Anatomy, College of Dentistry, Pusan National University)
  • 손장호 (울산대학교 의과대학 울산대학교병원 치과/구강악안면외과) ;
  • 조영철 (울산동강병원 치과/구강악안면외과) ;
  • 류성호 (울산대학교 의과대학 울산대학교병원 치과/구강악안면외과) ;
  • 김규천 (부산대학교 치과대학 치의학과 해부학교실) ;
  • 성일용 (울산대학교 의과대학 울산대학교병원 치과/구강악안면외과) ;
  • 박봉수 (부산대학교 치과대학 치의학과 해부학교실)
  • Published : 2004.12.31
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Abstract

Photodynamic therapy(PDT) has advanced to clinical trials for the treatment of a variety of solid tumors and presents an alternative treatment option for tumors resistant to chemo-and/or radio-therapy. PDT is based on the combination of laser light of appropriate wavelength and energy to activate a systemically or locally applied photosensitizer that concentrates preferentially in malignant tissues. In this study, phototoxicity of laser EIT 21 was analysed in human osteosarcoma cell(HOS) and the second objective of this study was to determine the ability of laser EIT 21 to induce apoptosis. This study demonstrated that laser EIT 21 had a phototoxicity to HOS cells. In order to examinate whether cell death was induced by necrosis or apoptosis, variety of techniques which assess apoptosis were used. TUNEL assay showed only a few the positive reaction on condensed nuclei. It is hard to find condensed or fragmented nuclei on HOS cells irradiated with laser EIT 21 in Hemastat and AO/EB stain. By DNA electrophoresis, cells also did not show DNA degradation characteristic of apoptosis with a ladder pattern of DNA fragments. Apoptosis-related factors were analyzed by western blotting. The expression of p53 was constant and cells irradiated with laser did not show the caspase-3 and PARP degradation, therefore we suggest that p53 and caspase-3 are not involved in laserinduced cell death.

광역동치료(PDT)는 갖가지 고형 종양의 치료를 위한 임상적시도로 시작되어 화학요법 및 방사선 요법에 내성이 있는 종양의 대체 치료법의 하나로 제시되고 있다. PDT는 전신적 또는 국소적으로 투여하여 유해한 조직에 선택적으로 농축되도록 한 광증감제와 이를 활성화하는 적절한 파장과 에너지를 가진 레이저광의 조합에 기초한다. 이 연구에서는, 인체 골육종 세포(HOS)에 미치는 레이저 EIT 21의 광독성 효과에 대해 알아보았으며, 그런 광독성 효과가 세포고사를 유발하는가를 규명하고자 하였다. 본 연구는 레이저 EIT 21이 HOS 세포에 대해 광독성을 효과를 가진다는 것을 증명했다. 세포 죽음이 세포괴사에 의해 유발되는지 아니면 세포고사에 의해 유발되는지를 알아보기 위해 세포 고사를 평가 하는 여러 실험 기법을 이용하였다. TUNEL 분석은 극소수만이 응축된 핵의 양성반응을 보여주었다. Hemacolor와 AO/EB 염색 또한 대부분의 세포가 괴사로 죽는 것을 보여주었다. 레이저 EIT로 조사된 HOS 세포에서 응축되거나 분절된 핵을 발견하는 것은 어려웠다. DNA 전기영동에서, 세포고사에서 보여지는 DNA 분절의 전형적인 특징인 사다리형 절편 형태(ladder fragmentation pattern)가 나타나지 않았다. Western blotting에 의한 분석에서 p53의 발현은 일정하게 나타났고 레이저로 조사된 세포는 caspase-3과 PARP의 분열을 나타내지 않는 것으로 보아 레이저 유도 세포 죽음(laser-induced cell death)은 p53과는 관련이 없는 것 같다.

Keywords

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