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Year:
1999
| Volume: 7
| Issue: 3
| Pages: 91-95
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Original Article |
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SPONTANEOUS AND RADIATION-INDUCED CELL DEATH IN HeLa S3 HUMAN CARCINOMA
Bozidarka ZARIC, Bratoljub H. MILOSAVLJEVIC, Marija RADOJCIC |
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DOI:
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Abstract: |
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Background: Radiation biologists have classified radiation-induced cell death based on cell proliferative capacity to either mitotic or interphase death. Cytologists have revealed two morphologically and biochemically diverse forms of cell death, apoptosis and necrosis. While the knowledge of the former is already well exploited by radiologists, cell susceptibility to apoptosis or necrosis is still under investigation. We studied characteristics of spontaneous cell death, dose dependence and time course of radiation-induced cell death of human uterine cervix epitheloid carcinoma HeLa S3 in culture. Methods: Cells were irradiated with 2-40 Gy of g-rays. The effects on growth, viability, morphology and genomic DNA structure were followed 24-72 h after irradiation. Cell viability was evaluated by trypan blue exclusion assay and cell morphology by in situ DNA staining with propidium iodide. DNA fragmentation pattern was determined by electrophoresis on 2% agarose gels. Results: At all cell densities, 15-20% of cells were PI positive and their DNA was fragmented to a high molecular size (>=20kb), but not to internucleosomal ladder. A significant decrease in viability to 35% was observed 72 h post 40 Gy irradiation. It corresponded to 55% of PI positive cells. A smear of smaller DNA fragments (0.1-1 kb), 24 h after 10-20 Gy irradiation was considered as proof that the dominant form of radiation-induced cell death was necrosis. Conclusion: The dominant form of radiation-induced cell death in HeLa S3 population is necrosis and the radiation dose which caused 50% of death after 72 h (termed ND50) was between 30-40 Gy. |
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Key words:
Cell death; Ionizing radiation; Human uterine carcinoma |
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