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 β-ACTIN, β-MICROGLOBULIN, Cu-Zn SUPEROXIDE DISMUTASE (SOD-1) and CATALASE. In cells exposed to 3·6 Gy low dose-rate γ-irradiation we observed increased levels of the GADD45 transcript and lower transcript levels of the genes TOPOISOMERASE IIα, FACC, CYCLIN A and CYCLIN B. No differences were detected in the transcript levels of β-ACTIN, β-MICROGLOBULIN, SOD-1, URACYL-DNA GLYCOSYLASE, CYCLIN C, CYCLIN E, CYCLIN D1, CYCLIN D2, CYCLIN D3, TOPOISOMERASE I and TOPOISOMERASE IIβ.