Comet assay slides 에서 나타난 apoptosis 평가에서 함수 및 탈수 겔의 비교

A Comparison of Hydrated versus Dehydrated Gels for Evaluation of Apoptosis in Comet Assay Slides

  • 최민철 (경상대학교 수의과대학 및 동물의학 연구소) ;
  • 수즌엠러루 (미국 콜로라도 주립대학 방사선 및 방사선 생물학 교실) ;
  • 에드워드엘질럿 (미국 콜로라도 주립대학 방사선 및 방사선 생물학 교실)
  • 발행 : 1996.06.01

초록

Comet assay 는 포유류 세포에서 DNA의 파괴를 측정하는데 있어 신속하고 단순하며 시각적이고 민감한 방법이다. Apoptosis에서는 세포핵의 광범위한 DNA의 붕괴가 일어나므로 comet assay는 종양세포에서 apoptosis가 발생되었는가를 알아내는데 유용하다. 본 연구는 apoptosis 연구의 결과가 변화되지 않도록 comet assay slides를 좀 더 오래 보관할 수 있는 방법을 개발하고자 시행되었다. 개의 종양세포를 가지고 alkali comet assay를 끝낸 뒤 slides를 진공 건조기에서 꺼내서 증류수로 점적하여 10-20분간 침수시키고 현광현미경하에서 육안적으로 관찰하였다. 건조후 3-4일, 1주, 2주, 3주, 4주 및 7주의 slides에서 apoptosis 회복율(%)은 각각 98.1, 98.3, 99.4, 80.8 및 35.2%이었다. 3주 이내의 slides에서 대조군과 비교하여 apoptosis 회복율에서 차이가 없었으나 4주 이상의 slides를 건조후 침수시키는 방법을 이용하였을 때 apoptosis 평가에서 건조 후 3주간까지는 처음의 결과와 차이가 없으며, 이 방법을 이용하여 comet slide의 좀 더 긴 기간이 보관과 보관후의 재평가에서 이용될 수 있는 좋은 방법이 된다.

키워드

참고문헌

  1. Mutation Res v.307 The effect of various antitoxidants and other modifying agents on oxygen-radical-generated DNA damage in human lympocytes in the comet assay Anderson D;Yu TW;Phillips BJ;Schmezer P
  2. Cancer Res v.53 Hypersensitivity of human lympocytes to UV-B and solar irradiation Arlett CF;Lowe JE;Harcourt SA(et al.)
  3. Radiat Res v.134 DNA double-strand break rejoining deficiency on TK6 and other human B-lymphoblast cell lines Evans HH;Ricaanti M;Horng MF(et al.)
  4. Mutation Res v.339 The comet assay: a comprehensive review Fairbairn DW;Olive PL;O'Neill KL
  5. Mutation Res v.273 UV-C sensitivity of unstimulated and stimualted human lympocytosis from normal and xeroderma pigmentosum donors in the comet assay: a potential diagnostic technique Green MHL;Lowe JE;Harcourt SA
  6. Lab Invest v.14 Vulnerability to oxygen radical is more important than imparied repair in hepatocytic deoxyribonucleic acid damage in aging Higami Y;Shimogawa I;Okimoto T;Ikeda T
  7. Carcinogenesis v.14 Genotoxic activity of 1-chloromethypyrene in stomach epithelium in vivo : intensity of the stomach scintillation UDS assay Kenelly JC;Lane MP;Barker JA(et al.)
  8. Mutation Res v.288 A European review Mckevery-Martin VJ;Green MHL;Schmezer P;Pool-Zobel BL;De Meo MP;Collins A
  9. Cell Prolif v.25 Comet assay Muller WU;Bauch T;Streffer M;Niedereichholz F;Boker W
  10. Cell Prolif v.25 Growth fraction measured using the comet assay Olive PL;Beneath JP
  11. Mutation Res v.294 Induction and rejoining of radiation-induced DNA single-strand breaks : "tail moment" as a function of position in the cell cycle Olive PL;Banath JP
  12. Radiat Res v.142 Radiation-induced DNA double-stand breaks produced in histone-depleted tumor cell nuclei measured using the neutral comet assay Olive PL;Banath JP
  13. Radat Res v.122 Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay Olive PL;Beneath JP;Durand RE
  14. Cytometry v.16 DNA strand breakage and DNA structure influence staining with propidium iodide using the alkaline comet assay Olive PL;Banath JP;Fjell CD
  15. Cancer Res v.54 Lack of a correlation between radiosensitivity and DNA double-strand break induction or rejoning in six human tumor cell lines Olive PL;Beneath JP;Macpail HS
  16. Cancer Res v.53 Gel electrophoresis of individual cells to quantify hypoxic fraction in human cancers Olive PL;Durand RE;Le Riche J;Olibvotto IA;Jaction SM
  17. Exp Cell Res v.198 Factors influencing DNA migration from individual cells subjected to gel electrophoresis Olive PL;Durand RE;Wlodek D;Banath JP
  18. Radiat Res v.136 Radiation-apoptosis measured in TK6 human B lymphoblast cells using the comet assay Olive PL;Frazer G;Banath JP
  19. Mutant Res v.319 Analysis of single-cell gel electrophoresis using laser-scanning microscopy O'Neill KL;Fairburun DW;Standing MD
  20. Int J Radiat Biol v.52 Belomycin, in contrast to gamma irradiation, induces extreme variation of DNA strand breakage from cell to cell $\"{O}$stling O;Johnson KJ
  21. Biochem Biophys Res Commun v.123 Microelectroploretic study of radiation-induced DNA damages in individual mammalian cells Ostling O;Johnson KJ
  22. Exp Cell Res v.175 A simple technique for quantitation of low levels of DNA damage in individual cells Singh NP;McCoy MT;Tice RR;Schnider EL
  23. Radiat Res v.136 Oxygen in human tumors: correlations between methods of measurement and response to therapy Stone HB;Brown JM;Phillips TL;Sutherland RM
  24. Radiat Res v.137 Radiosensitivity of CD 45RO+memory and CD45RO-native T cell in culture Uzawa A;Suzuki G;Nikata Y;Akashi M;Ohyama H;Akanuma A