Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry
Micronucleus assay is a test used to evaluate genotoxic damage in cells, which can be caused by various factors, like ionizing radiation. Interactions between radiation energies and DNA can cause breakage, leading to use chromosomal mutations or loss of genetic material, important events that could...
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Brazilian Radiation Protection Society (Sociedade Brasileira de Proteção Radiológica, SBPR)
2019-02-01
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| Series: | Brazilian Journal of Radiation Sciences |
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| Online Access: | https://bjrs.org.br/revista/index.php/REVISTA/article/view/572 |
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| author | Leticia Bonfim Luma Ramirez de Carvalho Daniel Perez Vieira |
| author_facet | Leticia Bonfim Luma Ramirez de Carvalho Daniel Perez Vieira |
| author_sort | Leticia Bonfim |
| collection | DOAJ |
| description | Micronucleus assay is a test used to evaluate genotoxic damage in cells, which can be caused by various factors, like ionizing radiation. Interactions between radiation energies and DNA can cause breakage, leading to use chromosomal mutations or loss of genetic material, important events that could be induced in solid tumors to mitigate its expansion within human body. Melanoma has been described as a tumor with increased radio resistance. This work evaluated micronuclei percentages (%MN) in human melanoma cells (SK-MEL-37), irradiated by gamma radiation, with doses between 0 and 16Gy. Cell suspensions were irradiated in PBS by a 60Co source in doses between 0 and 16Gy, and incubated by 48h. Then cell membranes were lysed in the presence of SYTOX Green and EMA dyes, preserving nuclear membranes. Using this method, EMA-stained nuclei could be discriminated as those derived from dead cells, and SYTOX nuclei and micronuclei could be quantified. Micronuclei percentages were found to be proportional to dose, (R2 = 0.997). Only the highest dose (16Gy) could induce statistically significant increase of MN (p<0.0001), although cultures irradiated by 4, 8 and 16Gy showed significant increase of dead cell fractions. Calculation of the nuclei-to-beads ratio showed that 8 and 16Gy could reduce melanoma cell proliferation. Results showed that although cell death and loss of proliferative capacity could be observed on cultures irradiated at lower doses, genotoxic damage could be induced only on a higher dose. Resistance to radiation-induced genotoxicity could explain a relatively high radio resistance of melanoma tumors. |
| format | Article |
| id | doaj-art-bbef431ee9464bebbe25869f6939082e |
| institution | Kabale University |
| issn | 2319-0612 |
| language | English |
| publishDate | 2019-02-01 |
| publisher | Brazilian Radiation Protection Society (Sociedade Brasileira de Proteção Radiológica, SBPR) |
| record_format | Article |
| series | Brazilian Journal of Radiation Sciences |
| spelling | doaj-art-bbef431ee9464bebbe25869f6939082e2025-08-20T03:27:36ZengBrazilian Radiation Protection Society (Sociedade Brasileira de Proteção Radiológica, SBPR)Brazilian Journal of Radiation Sciences2319-06122019-02-0172A (Suppl.)10.15392/bjrs.v7i2A.572434Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometryLeticia Bonfim0Luma Ramirez de Carvalho1Daniel Perez Vieira2Institute of Energy and Nuclear ResearchInstituto de Pesquisas Energéticas e Nucleares (IPEN / CNEN - SP) Av. Professor Lineu Prestes 2242 CEP 05508-000 São Paulo, SPInstituto de Pesquisas Energéticas e Nucleares (IPEN / CNEN - SP) Av. Professor Lineu Prestes 2242 CEP 05508-000 São Paulo, SPMicronucleus assay is a test used to evaluate genotoxic damage in cells, which can be caused by various factors, like ionizing radiation. Interactions between radiation energies and DNA can cause breakage, leading to use chromosomal mutations or loss of genetic material, important events that could be induced in solid tumors to mitigate its expansion within human body. Melanoma has been described as a tumor with increased radio resistance. This work evaluated micronuclei percentages (%MN) in human melanoma cells (SK-MEL-37), irradiated by gamma radiation, with doses between 0 and 16Gy. Cell suspensions were irradiated in PBS by a 60Co source in doses between 0 and 16Gy, and incubated by 48h. Then cell membranes were lysed in the presence of SYTOX Green and EMA dyes, preserving nuclear membranes. Using this method, EMA-stained nuclei could be discriminated as those derived from dead cells, and SYTOX nuclei and micronuclei could be quantified. Micronuclei percentages were found to be proportional to dose, (R2 = 0.997). Only the highest dose (16Gy) could induce statistically significant increase of MN (p<0.0001), although cultures irradiated by 4, 8 and 16Gy showed significant increase of dead cell fractions. Calculation of the nuclei-to-beads ratio showed that 8 and 16Gy could reduce melanoma cell proliferation. Results showed that although cell death and loss of proliferative capacity could be observed on cultures irradiated at lower doses, genotoxic damage could be induced only on a higher dose. Resistance to radiation-induced genotoxicity could explain a relatively high radio resistance of melanoma tumors.https://bjrs.org.br/revista/index.php/REVISTA/article/view/572micronucleus assaymelanomaradiationgenotoxic damage.. |
| spellingShingle | Leticia Bonfim Luma Ramirez de Carvalho Daniel Perez Vieira Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry Brazilian Journal of Radiation Sciences micronucleus assay melanoma radiation genotoxic damage.. |
| title | Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry |
| title_full | Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry |
| title_fullStr | Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry |
| title_full_unstemmed | Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry |
| title_short | Evaluation of radiation-induced genotoxicity on hu-man melanoma cells (SK-MEL-37) by flow cytometry |
| title_sort | evaluation of radiation induced genotoxicity on hu man melanoma cells sk mel 37 by flow cytometry |
| topic | micronucleus assay melanoma radiation genotoxic damage.. |
| url | https://bjrs.org.br/revista/index.php/REVISTA/article/view/572 |
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