TY - CONF
T1 - Irradiation of extracellular matrix proteins affects their molecular structure and mediates epithelial cell behavior
T2 - Molecular Oncology Volume 18: Supplement: EACR 2024: Innovative Cancer Science, 10-13 June 2024, Rotterdam, Netherlands
AU - Meshram, Yamini
AU - Cartmell, Sarah
AU - Kirwan, Cliona
AU - Tuieng, Ren Jie
AU - Sherratt, Michael
PY - 2024/6/6
Y1 - 2024/6/6
N2 - IntroductionRadiotherapy is an important treatment for breast cancertreatment, but exposure of healthy tissues to therapeuticX-ray doses is associated with side effects includingsecondary cancers and fibrosis. Whilst the impact of X-ray exposure on cells is well characterized, theinteractions between ionizing radiation and thesurrounding extracellular matrix are poorly defined. Thisstudy aimed to use a model extracellular matrix (ECM)system Matrigel® to characterize the impact oftherapeutic X-ray doses on ECM structure anddownstream cell behavior.Material and MethodsMatrigel®, a reconstituted basement membrane matrixderived from Engelbreth-Holm-Swarm (EHS) mousesarcoma was exposed to X-ray radiation (doses of 50Gyor 100Gy). The impact of X-ray exposure on Matrigel®composition, protein abundance was assessed by massspectrometry (LC-MS/MS). Irradiation induced changesin protein structure were identified by peptide locationfingerprinting. The influence of irradiated Matrigel® onthe behavior of immortalized mouse mammary epithelialcells (EpH4) was assessed by live cell imaging (celladherence, viability, and proliferation) and bytranscriptomic analysis of extracted mRNA.Results and DiscussionsExposure to therapeutic X-ray doses (50 and 100 Gy) hadno effect Matrigel® protein identity or relativeabundance but did induce significant changes in localizedtryptic peptide yield for the basement membrane proteinslaminin, collagen IV, nidogen, and heparan sulfateproteoglycans. Epithelial cell adhesion was significantlyreduced on irradiated Matrigel® (at both X-ray doses).Hierarchical clustering of the top 50 differentiallyexpressed genes identified both dose dependentupregulation (17 genes) and downregulation (33 genes)following exposure of cells to 100Gy irradiatedMatrigel®. Significantly affected GO processes includedcell growth and shape and proliferation.ConclusionThis study demonstrates that exposure of ECM proteinsto therapeutic X-ray doses can significantly alter proteinstructure and in the behavior of adhered cells. Weconclude that therapeutic X-ray regimes have thepotential to influence normal tissue function as aconsequence of cell/matrix interactions.
AB - IntroductionRadiotherapy is an important treatment for breast cancertreatment, but exposure of healthy tissues to therapeuticX-ray doses is associated with side effects includingsecondary cancers and fibrosis. Whilst the impact of X-ray exposure on cells is well characterized, theinteractions between ionizing radiation and thesurrounding extracellular matrix are poorly defined. Thisstudy aimed to use a model extracellular matrix (ECM)system Matrigel® to characterize the impact oftherapeutic X-ray doses on ECM structure anddownstream cell behavior.Material and MethodsMatrigel®, a reconstituted basement membrane matrixderived from Engelbreth-Holm-Swarm (EHS) mousesarcoma was exposed to X-ray radiation (doses of 50Gyor 100Gy). The impact of X-ray exposure on Matrigel®composition, protein abundance was assessed by massspectrometry (LC-MS/MS). Irradiation induced changesin protein structure were identified by peptide locationfingerprinting. The influence of irradiated Matrigel® onthe behavior of immortalized mouse mammary epithelialcells (EpH4) was assessed by live cell imaging (celladherence, viability, and proliferation) and bytranscriptomic analysis of extracted mRNA.Results and DiscussionsExposure to therapeutic X-ray doses (50 and 100 Gy) hadno effect Matrigel® protein identity or relativeabundance but did induce significant changes in localizedtryptic peptide yield for the basement membrane proteinslaminin, collagen IV, nidogen, and heparan sulfateproteoglycans. Epithelial cell adhesion was significantlyreduced on irradiated Matrigel® (at both X-ray doses).Hierarchical clustering of the top 50 differentiallyexpressed genes identified both dose dependentupregulation (17 genes) and downregulation (33 genes)following exposure of cells to 100Gy irradiatedMatrigel®. Significantly affected GO processes includedcell growth and shape and proliferation.ConclusionThis study demonstrates that exposure of ECM proteinsto therapeutic X-ray doses can significantly alter proteinstructure and in the behavior of adhered cells. Weconclude that therapeutic X-ray regimes have thepotential to influence normal tissue function as aconsequence of cell/matrix interactions.
U2 - 10.1002/1878-0261.13683
DO - 10.1002/1878-0261.13683
M3 - Abstract
SP - 334
EP - 335
ER -