Abstract
This work examined the importance of radiation-induced and ligand-induced EGFR-ERK signaling for the regulation of DNA repair proteins XRCC1 and ERCC1 in prostate carcinoma cells, DU145 (TP53(mut)), displaying EGFR-TGFA-dependent autocrine growth and high MAPK (ERK1/2) activity, and LNCaP (TP53(wt)) cells expressing low constitutive levels of ERK1/2 activity. Using quantitative RT-PCR and Western analyses, we determined that ionizing radiation activated the DNA repair genes XRCC1 and ERCC1 in an ERK1/2-dependent fashion for each cell line. After irradiation, a rapid increase followed by a decrease in ERK1/2 activity preceded the increase in XRCC1/ERCC1 expression in DU145 cells, while only the rapid decrease in ERK1/2 preceded the increase in XRCC1/ERCC1 expression in LNCaP cells. Administration of EGF, however, markedly increased the up-regulation of phospho-ERK, ERCC1 and XRCC1 in both cell lines. Although the EGFR inhibitor tyrphostin (AG-1478) and the MEK inhibitor PD90859 both attenuated EGF-induced levels of the ERCC1 and XRCC1 protein, PD98059 blocked the induction of ERCC1 and XRCC1 by radiation more effectively in both cell lines. Inhibition of ERK at a level that reduced the up-regulation of DNA repair led to the persistence of apurinic/apyrimidinic (AP) sites of DNA damage and increased cell killing. Taken together, these data imply a complex control of DNA repair activation that may be more generally dependent on MAPK (ERK1/2) signaling than was previously noted. These data provide novel insights into the capacity of the EGFR-ERK signaling to modulate DNA repair in cancer cells and into the functional significance of this signaling.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenocarcinoma / pathology*
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Autocrine Communication
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Cobalt Radioisotopes
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DNA Damage
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DNA Repair / drug effects
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DNA Repair / physiology*
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DNA Repair / radiation effects
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DNA-Binding Proteins / biosynthesis*
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DNA-Binding Proteins / genetics
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Endonucleases*
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Enzyme Inhibitors / pharmacology
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Epidermal Growth Factor / pharmacology*
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ErbB Receptors / drug effects
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ErbB Receptors / physiology*
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Flavonoids / pharmacology
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Gamma Rays*
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Gene Expression Regulation, Neoplastic* / drug effects
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Gene Expression Regulation, Neoplastic* / radiation effects
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Humans
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MAP Kinase Signaling System / drug effects
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MAP Kinase Signaling System / physiology*
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MAP Kinase Signaling System / radiation effects
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Male
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Mitogen-Activated Protein Kinase 1 / metabolism*
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Mitogen-Activated Protein Kinase 3
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Mitogen-Activated Protein Kinases / metabolism*
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Neoplasm Proteins / biosynthesis
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Neoplasm Proteins / drug effects
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Neoplasm Proteins / genetics
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Neoplasm Proteins / physiology*
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Prostatic Neoplasms / pathology*
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Protein Biosynthesis*
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Proteins / genetics
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Quinazolines
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Tumor Cells, Cultured / drug effects
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Tumor Cells, Cultured / metabolism
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Tumor Cells, Cultured / radiation effects
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Tyrphostins / pharmacology
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X-ray Repair Cross Complementing Protein 1
Substances
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Cobalt Radioisotopes
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DNA-Binding Proteins
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Enzyme Inhibitors
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Flavonoids
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Neoplasm Proteins
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Proteins
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Quinazolines
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Tyrphostins
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X-ray Repair Cross Complementing Protein 1
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XRCC1 protein, human
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RTKI cpd
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Epidermal Growth Factor
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ErbB Receptors
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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Mitogen-Activated Protein Kinases
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ERCC1 protein, human
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Endonucleases
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2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one