title: Impairment of the DNA repair and growth arrest pathways by p53R2 silencing enhances DNA damage-induced apoptosis in a p53-dependent manner in prostate cancer cells.
abstract: p53R2 is a p53-inducible ribonucleotide reductase that contributes to DNA repair by supplying deoxynucleotide triphosphate pools in response to DNA damage. In this study, we found that p53R2 was overexpressed in prostate tumor cell lines compared with immortalized prostatic epithelial cells and that the protein was induced upon DNA damage. We investigated the effects of p53R2 silencing on DNA damage in LNCaP cells (wild-type p53). Silencing p53R2 potentiated the apoptotic effects of ionizing radiation and doxorubicin treatment as shown by increased sub-G(1) content and decreased colony formation. This sensitizing effect was specific to DNA-damaging agents. Comet assay and gamma-H2AX phosphorylation status showed that the decreased p53R2 levels inhibited DNA repair. Silencing p53R2 also reduced the levels of p21(WAF1/CIP1) at the posttranscriptional level, suggesting links between the p53-dependent DNA repair and cell cycle arrest pathways. Using LNCaP sublines stably expressing dominant-negative mutant p53, we found that the sensitizing effect of p53R2 silencing is mediated by p53-dependent apoptosis pathways. In the LNCaP sublines (R273H, R248W, and G245S) that have defects in inducing p53-dependent apoptosis, p53R2 silencing did not potentiate DNA damage-induced apoptosis, whereas p53R2 silencing was effective in a LNCaP subline (P151S) which retains the ability to induce p53-dependent apoptosis. This study shows that p53R2 is a potential therapeutic target that could be used to enhance the effectiveness of ionizing radiation or DNA-damaging chemotherapy in a subset of patients with prostate cancer.
authors.affiliation: Department of Internal Medicine, University of California, Davis, School of Medicine, 4860 Y Street, Suite 3500, Sacramento, CA 95817, USA.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Devlin
authors.forename: Phillip C
authors.initials: PC
authors.lastname: Mack
authors.forename: Rebekah A
authors.initials: RA
authors.lastname: Burich
authors.forename: Paul H
authors.initials: PH
authors.lastname: Gumerlock
authors.forename: Hsing-Jien
authors.initials: HJ
authors.lastname: Kung
authors.forename: Maria
authors.initials: M
authors.lastname: Mudryj
authors.forename: Ralph W
authors.initials: RW
authors.lastname: deVere White
mesh_terms: D017209:Apoptosis; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000737:chemistry*; D045744:Cell Line, Tumor; D004249:DNA Damage; D004260:DNA Repair; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D005799:Genes, Dominant; D016158:Genes, p53; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D012264:Ribonucleotide Reductases / Q000737:chemistry*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D034741:RNA, Small Interfering; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C405947:RRM2B protein, human; D012264:Ribonucleotide Reductases