Evaluation of radiation-induced genotoxicity on human melanoma cells (SK-MEL-37) by flow cytometry
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , |
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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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. DE:12133:810:NVIEIRA, DANIEL P.:3158:810:NORIGINAL24108.pdf24108.pdfapplication/pdf412269http://repositorio.ipen.br/bitstream/123456789/28283/1/24108.pdf99b4df151890b5555d87bc6ad44b3215MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ipen.br/bitstream/123456789/28283/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52123456789/282832022-03-24 14:10:11.009oai:repositorio.ipen.br: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Repositório InstitucionalPUBhttp://repositorio.ipen.br/oai/requestbibl@ipen.bropendoar:45102022-03-24T14:10:11Repositório Institucional do IPEN - Instituto de Pesquisas Energéticas e Nucleares (IPEN)false |
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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info:eu-repo/semantics/conferenceObject |
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conferenceObject |
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http://repositorio.ipen.br/handle/123456789/28283 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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I |
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Associa????o Brasileira de Energia Nuclear |
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Associa????o Brasileira de Energia Nuclear |
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