Characterization of the role of Fhit in maintenance of genomic integrity following low dose radiation, in vivo and in vitro

Ya Wang

关键词: CARCINOGENESIS;    CARCINOGENS;    GENE MUTATIONS;    GENES;    GENETICS;    HYPOTHESIS;    IN VITRO;    IN VIVO;    INSTABILITY;    IONIZING RADIATIONS;    MAINTENANCE;    MICE;    MUTAGENESIS;    MUTANTS;    NEOPLASMS;    PROTEINS;    REPAIR;    ST;   

DOI  :  10.2172/990617
RP-ID  :  DOE/SC0002539
PID  :  OSTI ID: 990617
Others  :  TRN: US201206%%350

学科分类：生物科学（综合）

美国|英语

来源: SciTech Connect

【 摘 要 】

The major goal of this study is to determine the effects of the Fhit pathway on low dose ({le} 0.1 Gy) ionizing radiation (IR)-induced genetic instability. Reduction of Fhit protein expression is observed in most solid tumors particularly in those tumors resulting from exposure to environmental carcinogens. Therefore, characterization of the role of the Fhit-dependent pathway in preventing low dose IR-induced genetic instability will provide useful parameters for evaluating the low dose IR-induced risk of mutagenesis and carcinogenesis. We pursued 3 specific aims to study our hypothesis that the Fhit-dependent pathways maintain genomic integrity through adjusting checkpoint response and repair genes expression following low dose IR. Aim 1: Determine whether Fhit interaction with RPA is necessary for Fhit to affect the cellular response to low dose IR. We combined the approaches of in vitro (GST pull-down and site-directed mutagenesis) and in vivo (observing the co-localization and immunoprecipitation of Fhit and RPA in Fhit knock out mouse cells transfected with mutant Fhit which has lost ability to interact with RPA in vitro). Aim 2: Determine the role of genes whose expression is affected by Fhit in low dose irradiated cells. We analyzed the distinct signature of gene expression in low dose irradiated Fhit-/- cells compared with Fhit+/+ cells by combining microarray, gene transfection and siRNA approaches. Aim 3: Determine the role of Fhit in genetic susceptibility to low dose IR in vivo. We compared the gene mutation frequency and the fragile site stability in the cells isolated from the Fhit+/+ and Fhit-/- mice at 1.5 years following low dose IR. These results determine the role of the Fhit-dependent pathway in maintaining genomic integrity in vitro and in vivo, which provide a basis for choosing surrogate markers in the Fhit-dependent pathway to evaluate low dose IR-induced risk of mutagenesis and carcinogenesis.