DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.

Our genome is constantly exposed to both endogenous and exogenous agents that cause genetic damage. Among the endogenous agents we can find reactive oxygen species (ROS) as cellular metabolism or immune response products (cell to cell genotoxicity). The cellular response to these damages is to repai...

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Autores:
Camargo Guerrero, Mauricio
Tipo de recurso:
Investigation report
Fecha de publicación:
2007
Institución:
Ministerio de Ciencia, Tecnología e Innovación
Repositorio:
Repositorio Minciencias
Idioma:
OAI Identifier:
oai:repositorio.minciencias.gov.co:20.500.14143/38054
Acceso en línea:
https://colciencias.metadirectorio.org/handle/11146/38054
http://colciencias.metabiblioteca.com.co
Palabra clave:
Apoptosis
OGG1
BER
DNA Repair
Polymorphism
ROS
XPD
XRCC1
Cell to cell genotoxicity
Rights
openAccess
License
http://purl.org/coar/access_right/c_abf2
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dc.title.spa.fl_str_mv DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
title DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
spellingShingle DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
Apoptosis
OGG1
BER
DNA Repair
Polymorphism
ROS
XPD
XRCC1
Cell to cell genotoxicity
title_short DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
title_full DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
title_fullStr DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
title_full_unstemmed DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
title_sort DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.
dc.creator.fl_str_mv Camargo Guerrero, Mauricio
dc.contributor.author.none.fl_str_mv Camargo Guerrero, Mauricio
dc.contributor.corporatename.spa.fl_str_mv Universidad de Antioquia (Medellín, Colombia)
dc.contributor.researchgroup.none.fl_str_mv Inmunodeficiencia primarias
dc.subject.proposal.spa.fl_str_mv Apoptosis
OGG1
BER
DNA Repair
Polymorphism
ROS
XPD
XRCC1
Cell to cell genotoxicity
topic Apoptosis
OGG1
BER
DNA Repair
Polymorphism
ROS
XPD
XRCC1
Cell to cell genotoxicity
description Our genome is constantly exposed to both endogenous and exogenous agents that cause genetic damage. Among the endogenous agents we can find reactive oxygen species (ROS) as cellular metabolism or immune response products (cell to cell genotoxicity). The cellular response to these damages is to repair them or to undergo programmed cell death. DNA repair mechanisms are part of a vital defence system designed to protect the integrity of the genome. Deficiencies in this system most probably lead to cell death or cancer development. There is a wide variability in coding regions of DNA repair genes which may lead to an increased or decrease in repair capability. There is increasing evidence suggesting that a decrease in DNA repair capacity is a consequence of genetic variability and affects cancer susceptibility. Because of all this, the question of interest in our case would be as follows: ¿Does genetic variation in XRCC1 and OGG1 genes affect repair rate and apoptosis induced by endogenous ROS? Our hypothesis is that respiratory burst induces oxidative DNA damage and that this damage activates the base excision repair pathway (BER) and/or apoptosis, therefore, polymorphisms in genes involved in this pathway (XRCC1, OGG1) may affect the inter-individual repair rate of the damage induced by ROS and consequently affect the apoptotic process. In order to correlate these parameters we will determine the allelic frequency of two polymorphisms on gene XRCC1 (Arg194Trp and Arg399Gln), one polymorphism on gene OGG1 (Ser326Cys), and one polymorphism on gene XPD (Lys751Gln) by means of PCR-RFLP in a population of 174 healthy adults belonging to the university community. Secondly, we will obtain two cellular populations (mononuclear and polymorphonuclear cells) from peripheral blood and they will be co-cultured in order to determine repair rates and apoptosis of individuals that bear genetic polymorphisms by means of the Comet Assay and flow cytometry respectively. With this project we expect to establish a relation between polymorphisms in DNA repair genes and both repair and apoptosis events induced by ROS produced by polymorphonuclear (PMN) cells that undergo respiratory burst. According to the results we may be able to reach a genotype-phenotype relation that may allow us to establish the individual susceptibility to ROS-induced cell to cell genotoxicity and consequently to the development of preneoplasic or neoplasic processes; and last but not least, we expect to contribute to the identification of preventive therapeutic intervention targets, once determined which genotypes are more or less efficient in repairing genetic damage induced by endogenous ROS (cell to cell genotoxicity). Resumen Nuestro genoma esta constantemente expuesto a agentes causantes de daño genético tanto endógena como exógenamente. Entre los agentes endógenos se encuentran las especies reactivas de oxígeno como productos del metabolismo celular ó de la respuesta inmune (genotoxicidad célula-célula). La respuesta celular a estos daños es repararlos ó sufrir muerte celular programada. Los mecanismos de reparación del DNA componen un sistema vital de defensa diseñado para proteger la integridad del genoma. Deficiencias en este sistema muy probablemente conducen a la muerte celular o al desarrollo de cáncer. Existe una extensa variabilidad en las regiones codificantes de los genes de reparación del DNA que muy probablemente se traducen en un efecto ya sea de disminución o aumento en la capacidad de reparación. Hay evidencia creciente de que una reducción en la capacidad de reparación del DNA es una consecuencia de la variación genética y afecta la predisposición a cáncer. Con base en lo anterior se plantea la siguiente pregunta: ¿Afecta la variación genética en el gen XRCC1 y OGG1 la tasa de reparación y la apoptosis inducidas por ROS endógenos? Nuestra hipótesis es que la explosión respiratoria induce daño oxidativo en el DNA.
publishDate 2007
dc.date.issued.none.fl_str_mv 2007
dc.date.accessioned.none.fl_str_mv 2020-03-10T23:20:47Z
2020-12-17T22:15:09Z
dc.date.available.none.fl_str_mv 2020-03-10T23:20:47Z
2020-12-17T22:15:09Z
dc.type.spa.fl_str_mv Informe de investigación
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dc.format.extent.spa.fl_str_mv 80 páginas.
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spelling Camargo Guerrero, Mauricio105716a8ebf874820143f7a0163edd4d-1Universidad de Antioquia (Medellín, Colombia)Inmunodeficiencia primarias2020-03-10T23:20:47Z2020-12-17T22:15:09Z2020-03-10T23:20:47Z2020-12-17T22:15:09Z2007https://colciencias.metadirectorio.org/handle/11146/38054ColcienciasRepositorio Colcienciashttp://colciencias.metabiblioteca.com.coOur genome is constantly exposed to both endogenous and exogenous agents that cause genetic damage. Among the endogenous agents we can find reactive oxygen species (ROS) as cellular metabolism or immune response products (cell to cell genotoxicity). The cellular response to these damages is to repair them or to undergo programmed cell death. DNA repair mechanisms are part of a vital defence system designed to protect the integrity of the genome. Deficiencies in this system most probably lead to cell death or cancer development. There is a wide variability in coding regions of DNA repair genes which may lead to an increased or decrease in repair capability. There is increasing evidence suggesting that a decrease in DNA repair capacity is a consequence of genetic variability and affects cancer susceptibility. Because of all this, the question of interest in our case would be as follows: ¿Does genetic variation in XRCC1 and OGG1 genes affect repair rate and apoptosis induced by endogenous ROS? Our hypothesis is that respiratory burst induces oxidative DNA damage and that this damage activates the base excision repair pathway (BER) and/or apoptosis, therefore, polymorphisms in genes involved in this pathway (XRCC1, OGG1) may affect the inter-individual repair rate of the damage induced by ROS and consequently affect the apoptotic process. In order to correlate these parameters we will determine the allelic frequency of two polymorphisms on gene XRCC1 (Arg194Trp and Arg399Gln), one polymorphism on gene OGG1 (Ser326Cys), and one polymorphism on gene XPD (Lys751Gln) by means of PCR-RFLP in a population of 174 healthy adults belonging to the university community. Secondly, we will obtain two cellular populations (mononuclear and polymorphonuclear cells) from peripheral blood and they will be co-cultured in order to determine repair rates and apoptosis of individuals that bear genetic polymorphisms by means of the Comet Assay and flow cytometry respectively. With this project we expect to establish a relation between polymorphisms in DNA repair genes and both repair and apoptosis events induced by ROS produced by polymorphonuclear (PMN) cells that undergo respiratory burst. According to the results we may be able to reach a genotype-phenotype relation that may allow us to establish the individual susceptibility to ROS-induced cell to cell genotoxicity and consequently to the development of preneoplasic or neoplasic processes; and last but not least, we expect to contribute to the identification of preventive therapeutic intervention targets, once determined which genotypes are more or less efficient in repairing genetic damage induced by endogenous ROS (cell to cell genotoxicity). Resumen Nuestro genoma esta constantemente expuesto a agentes causantes de daño genético tanto endógena como exógenamente. Entre los agentes endógenos se encuentran las especies reactivas de oxígeno como productos del metabolismo celular ó de la respuesta inmune (genotoxicidad célula-célula). La respuesta celular a estos daños es repararlos ó sufrir muerte celular programada. Los mecanismos de reparación del DNA componen un sistema vital de defensa diseñado para proteger la integridad del genoma. Deficiencias en este sistema muy probablemente conducen a la muerte celular o al desarrollo de cáncer. Existe una extensa variabilidad en las regiones codificantes de los genes de reparación del DNA que muy probablemente se traducen en un efecto ya sea de disminución o aumento en la capacidad de reparación. Hay evidencia creciente de que una reducción en la capacidad de reparación del DNA es una consecuencia de la variación genética y afecta la predisposición a cáncer. Con base en lo anterior se plantea la siguiente pregunta: ¿Afecta la variación genética en el gen XRCC1 y OGG1 la tasa de reparación y la apoptosis inducidas por ROS endógenos? Nuestra hipótesis es que la explosión respiratoria induce daño oxidativo en el DNA.80 páginas.Informe;DNA Repair genes XRCC1 and OGG1 : Genotoxicity and apoptosis induced by ROS.Informe de investigaciónhttp://purl.org/coar/resource_type/c_18wshttp://purl.org/coar/resource_type/c_93fcTextinfo:eu-repo/semantics/reporthttps://purl.org/redcol/resource_type/PIDinfo:eu-repo/semantics/submittedVersionhttp://purl.org/coar/version/c_71e4c1898caa6e32info:eu-repo/semantics/submittedVersion2005-2007info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by/4.0/ApoptosisOGG1BERDNA RepairPolymorphismROSXPDXRCC1Cell to cell genotoxicityEstudiantes, Profesores, Comunidad científica colombiana, etc.11150517582208-2005Departamento Administrativo de Ciencia, Tecnología e Innovación [CO] ColcienciasPrograma Nacional en Ciencias BásicasTo evaluate the in vitro effects of polymorphisms in DNA repair genes XRCC1 (Arg194Trp and Arg399Gln) and OGG1 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ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.
d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:
i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.
ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, los consagrados por la SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.
e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, ACINPRO), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.
5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.
6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.
7. Término.
a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.
b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.
8. Varios.
a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.
b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.
c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.
d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.
