title: Use of semiquantitative reverse transcription-polymerase chain reaction to study gene expression in normal human skin fibroblasts following low dose-rate irradiation.
abstract: One way to study the effect of radiation on gene expression is to monitor changes in the levels of specific messenger RNAs. We describe the use of reverse transcription-polymerase chain reaction (RT-PCR) analysis, a faster and more sensitive procedure than the traditional techniques to monitor RNA levels. Using RT-PCR, we confirmed previous results showing increased levels of GADD45 transcripts after high dose-rate X-irradiation in normal human fibroblasts. No differences were observed in the transcript levels of beta-ACTIN, beta-MICROGLOBULIN, Cu-Zn SUPEROXIDE DISMUTASE (SOD-1) and CATALASE. In cells exposed to 3-6 Gy low dose-rate gamma-irradiation we observed increased levels of the GADD45 transcript and lower transcript levels of the genes TOPOISOMERASE II alpha, FACC, CYCLIN A and CYCLIN B. No differences were detected in the transcript levels of beta-ACTIN, beta-MICROGLOBULIN, SOD-1, URACYL-DNA GLYCOSYLASE, CYCLIN C, CYCLIN E, CYCLIN D1, CYCLIN D2, CYCLIN D3, TOPOISOMERASE I and TOPOISOMERASE II beta.
authors.affiliation: Radiation Biology and Health Physics Branch, AECL Research, ON, Canada.
authors.forename: S M
authors.initials: SM
authors.lastname: de Toledo
authors.forename: E I
authors.initials: EI
authors.lastname: Azzam
authors.forename: M K
authors.initials: MK
authors.lastname: Gasmann
authors.forename: R E
authors.initials: RE
authors.lastname: Mitchel
mesh_terms: D000199:Actins / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D016213:Cyclins / Q000235:genetics / Q000502:physiology; D004264:DNA Topoisomerases, Type I / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000235:genetics; D005347:Fibroblasts / Q000528:radiation effects; D015870:Gene Expression / Q000528:radiation effects*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D012867:Skin / Q000528:radiation effects
chemical_list: D000199:Actins; D016213:Cyclins; D012333:RNA, Messenger; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II