CYTOFLUOROMETRIC DETERMINATION OF EXCISION REPAIR OF UV-INDUCED PYRIMIDINE DIMERS IN EHRLICH ASCITIC CANCER CELLS
スポンサーリンク
概要
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The ability of excision repair of a single Ehrlich ascitic cancer cell was quantitated by cytofluorometry of unrepaired pyrimidine dimer-FITC complex per single nuclear Feulgen DNA. UV-irradiated Ehrlich ascitic cancer cells were made react with antibodies to denatured UV-DNA using the antisera which were absorbed with MBSA and heat denatured DNA. Then the immunofluorescent staining specific for pyrimidine dimers was combined with subsequent Feulgen nuclear reaction using the method reported in the previous paper (Fukuda et al., 1976). Then simultaneous cytofluorometric measurement of the amounts of pyrimidine dimer-FITC complex and nuclear Feulgen DNA was performed on a single tumor cell at various times of incubation after UV-irradiation. Before the occurrence of excision repair of UV-DNA lesions, the amount of pyrimidine dimers per unit amount of nuclear DNA was found equal in all tumor cells at various stages of cell cycle. After incubation for 3 hours, unrepaired pyrimidine dimers were largest in amount in polyploid cells and least in diploid cells; some diploid cells had no nuclear fluorescence of FITC with a faint fluorescence in the cytoplasm. It might indicate their completion of excision repair of UV-induced pyrimidine dimers followed by transport of them to the cytoplasm within the period. Thus the potency of excision repair of a cell in S phase which has not been demonstrated by the detection of unscheduled DNA synthesis could be quantitated using the immunofluorescent-Feulgen double staining. The order of the increase in the magnitude of excision repair in Ehrlich ascitic cancer cells at different stages of cell cycle was polyploid cells<G2 and S phase cells<diploid G1 cells. When non-absorbed antisera were used, a significant fluorescence was found in the nuclei of some unirradiated large Ehrlich ascitic cancer cells. The specific fluorescence was suspected to have some special relation to naturally occuring single-stranded DNA in the tumor cells.
- 日本組織細胞化学会の論文
著者
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Shima Akihiro
Laboratory of Radiation Biology, Department of Biological Sciences, School of Science, University of
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FUKUDA Masaru
Department of Pathology, Fukui Medical University
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Shima Akihiro
Laboratory Of Radiation Biology Faculty Of Science University Of Tokyo
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NAKANISHI KAZUO
Department of Pathology, Kyoto Prefectural University of Medicine
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FUJITA SETSUYA
Department of Pathology, Kyoto Prefectural University of Medicine
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Fujita Setsuya
Department Of Hygiene Gifu University School Of Medicine
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Fujita Setsuya
Dept. Of Ophthalmol. And Pathol. Kyoto Pref. Univ. Med.
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Fujita Setsuya
Louis Pasteur Center For Medical Research
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MUKAINAKA Teruo
Kyoto Prefectural University of Medicine
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MUKAINAKA Teruo
Department of Biochemistry, Kyoto Prefectural University of Medicine
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Fujita S
Louis Pasteur Center For Medical Research
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Fukuda Masaru
Department Of Pathology Kyoto Prefectural University Of Medicine
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Fukuda Masaru
Department Of Pathology Fukui Medical School
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Mukainaka Teruo
Department Of Pathology Kyoto Prefectural University Of Medicine
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Nakanishi K
Food Research And Development Laboratories Ajinomoto Co. Inc.
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Nakanishi Kazuo
Department Of Orthopaedic Surgery Okayama University Hospital Of Medicine And Dentistry
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Nakanishi Kazuo
Dept Of Pathology Kyoto Pref. Univ. Of Med.
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Shima Akihiro
Laboratory Of Radiation Biology Department Of Biological Sciences The University Of Tokyo
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Fujita Setsuya
Department Of Dynamic Pathology Research Institute For Neurological Diseases And Geriatrics Kyoto Pr
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Nakanishi Kazuo
Department Of Anesthesiology And Resuscitology Ehime University Graduate School Of Medicine
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Fukuda Masaru
Department Of Anatomy Kyorin University School Of Medicine
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Muranaka Takashi
Faculty Of Pharmaceutical Sciences Okayama University
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FUKUDA MASARU
Department of Pathology, Kyoto Prefectural University of Medicine
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