Cellular senescence, a phenomenon correlated to both aging and age-related pathologies, as well as interfering with tumor progression, is closely associated with gene expression changes based on transcriptional, posttranscriptional and epigenetic regulatory mechanisms, as also demonstrated by our group (Cell Death Differ. 2012, 19:713-21; FASEB J. 2014, 28:3720-33; PLoSOne 2014, 9:e98669). By applying the 2D-DIGE coupled to Mass Spectrometry analysis, we profiled quantitative proteomic changes occurring both in replicative and oxidative stress-induced senescence of human IMR90 fibroblasts. We found that twenty protein spots shifted their quantitative representation in the same direction (over- or down-expressed) in both conditions of senescence. Among the 25 sequence entries associated with these variations, we identified a group of 10 genes showing at least a two times decrease of the cognate mRNAs, indicating that their regulation during senescence occurs at transcriptional level. To study the functional significance of the down-regulation of these genes, we individually silenced them by RNA interference-mediated knockdown in young IMR90 cells. The results show that silencing of LEPRE1, LIMA1/EPLIN, MAGOHA or MAGOHB gene induces a senescent phenotype with the appearance of specific markers. We also demonstrated that the expression of these genes is down-regulated in other models of senescence such as in DNA damage-induced senescence or in WI-38 replicative and stress-induced senescence. Furthermore we showed that the under-expression of LEPRE1, LIMA1/EPLIN, MAGOHA or MAGOHB is due to modifications of the histone H3 methylation states. In addition we obtained preliminary evidences that declined expression of these genes is related to in vivo human cellular senescence. Finally, functional experiments are in progress to establish whether ectopic over-expression of the above genes could hamper the senescence process in IMR90 cells.
Proteomics unveils novel common genes mediating both replicative and stress-induced senescence / Comegna, Marika; Succoio, Mariangela; D’Ambrosio, C; Scaloni, A; Cimino, Filiberto; Faraonio, Raffaella. - (2015). (Intervento presentato al convegno 58° National Meeting of the Italian Society of Biochemistry and Molecular Biology ( Plenary Symposium “Biochemestry for an active, healty aging “) tenutosi a Urbino nel 14 th -16 th September 2015).
Proteomics unveils novel common genes mediating both replicative and stress-induced senescence.
COMEGNA, Marika;SUCCOIO, MARIANGELA;CIMINO, FILIBERTO;FARAONIO, RAFFAELLA
2015
Abstract
Cellular senescence, a phenomenon correlated to both aging and age-related pathologies, as well as interfering with tumor progression, is closely associated with gene expression changes based on transcriptional, posttranscriptional and epigenetic regulatory mechanisms, as also demonstrated by our group (Cell Death Differ. 2012, 19:713-21; FASEB J. 2014, 28:3720-33; PLoSOne 2014, 9:e98669). By applying the 2D-DIGE coupled to Mass Spectrometry analysis, we profiled quantitative proteomic changes occurring both in replicative and oxidative stress-induced senescence of human IMR90 fibroblasts. We found that twenty protein spots shifted their quantitative representation in the same direction (over- or down-expressed) in both conditions of senescence. Among the 25 sequence entries associated with these variations, we identified a group of 10 genes showing at least a two times decrease of the cognate mRNAs, indicating that their regulation during senescence occurs at transcriptional level. To study the functional significance of the down-regulation of these genes, we individually silenced them by RNA interference-mediated knockdown in young IMR90 cells. The results show that silencing of LEPRE1, LIMA1/EPLIN, MAGOHA or MAGOHB gene induces a senescent phenotype with the appearance of specific markers. We also demonstrated that the expression of these genes is down-regulated in other models of senescence such as in DNA damage-induced senescence or in WI-38 replicative and stress-induced senescence. Furthermore we showed that the under-expression of LEPRE1, LIMA1/EPLIN, MAGOHA or MAGOHB is due to modifications of the histone H3 methylation states. In addition we obtained preliminary evidences that declined expression of these genes is related to in vivo human cellular senescence. Finally, functional experiments are in progress to establish whether ectopic over-expression of the above genes could hamper the senescence process in IMR90 cells.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
