Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry

Detalhes bibliográficos
Autor(a) principal: BONFIM, LETICIA
Data de Publicação: 2018
Outros Autores: CARVALHO, LUMA R. de, VIEIRA, DANIEL P., INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
Tipo de documento: Artigo de conferência
Título da fonte: Repositório Institucional do IPEN
Texto Completo: http://repositorio.ipen.br/handle/123456789/28283
Resumo: 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.
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spelling 2018-01-11T11:07:14Z2018-01-11T11:07:14ZOctober 22-27, 2017http://repositorio.ipen.br/handle/123456789/28283Micronucleus 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.Submitted by Marco Antonio Oliveira da Silva (maosilva@ipen.br) on 2018-01-11T11:07:14Z No. of bitstreams: 1 24108.pdf: 412269 bytes, checksum: 99b4df151890b5555d87bc6ad44b3215 (MD5)Made available in DSpace on 2018-01-11T11:07:14Z (GMT). No. of bitstreams: 1 24108.pdf: 412269 bytes, checksum: 99b4df151890b5555d87bc6ad44b3215 (MD5)Associa????o Brasileira de Energia Nuclearanimal cellscell flow systemscell nucleicobalt 60lymphokinesmelanomasradiation dosesradiation effectsradiosensitivitysurvival curvesEvaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometryinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectINACIRio de Janeiro, RJBelo Horizonte, MGBONFIM, LETICIACARVALHO, LUMA R. deVIEIRA, DANIEL P.INTERNATIONAL NUCLEAR ATLANTIC CONFERENCEinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional do IPENinstname:Instituto de Pesquisas Energéticas e Nucleares (IPEN)instacron:IPEN241082017BONFIM, LETICIACARVALHO, LUMA R. deVIEIRA, DANIEL P.18-01Proceedings14185121333158BONFIM, LETICIA:14185:810:SCARVALHO, LUMA R. 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dc.title.pt_BR.fl_str_mv Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
title Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
spellingShingle Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
BONFIM, LETICIA
animal cells
cell flow systems
cell nuclei
cobalt 60
lymphokines
melanomas
radiation doses
radiation effects
radiosensitivity
survival curves
title_short Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
title_full Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
title_fullStr Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
title_full_unstemmed Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
title_sort Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
author BONFIM, LETICIA
author_facet BONFIM, LETICIA
CARVALHO, LUMA R. de
VIEIRA, DANIEL P.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
author_role author
author2 CARVALHO, LUMA R. de
VIEIRA, DANIEL P.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
author2_role author
author
author
dc.contributor.author.fl_str_mv BONFIM, LETICIA
CARVALHO, LUMA R. de
VIEIRA, DANIEL P.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
dc.subject.por.fl_str_mv animal cells
cell flow systems
cell nuclei
cobalt 60
lymphokines
melanomas
radiation doses
radiation effects
radiosensitivity
survival curves
topic animal cells
cell flow systems
cell nuclei
cobalt 60
lymphokines
melanomas
radiation doses
radiation effects
radiosensitivity
survival curves
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.
publishDate 2018
dc.date.evento.pt_BR.fl_str_mv October 22-27, 2017
dc.date.accessioned.fl_str_mv 2018-01-11T11:07:14Z
dc.date.available.fl_str_mv 2018-01-11T11:07:14Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.identifier.uri.fl_str_mv http://repositorio.ipen.br/handle/123456789/28283
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dc.publisher.none.fl_str_mv Associa????o Brasileira de Energia Nuclear
publisher.none.fl_str_mv Associa????o Brasileira de Energia Nuclear
dc.source.none.fl_str_mv reponame:Repositório Institucional do IPEN
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