USE OF SEMIQUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION TO STUDY GENE-EXPRESSION IN NORMAL HUMAN SKIN FIBROBLASTS FOLLOWING LOW DOSE-RATE IRRADIATION

One way to study the effect of radiation on gene expression is to monitor changes in the levels of specific messenger RNAs. Wee 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 shorting 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 DI, CYCLIN D2, CYCLIN D3, TOPOISOMERASES I and TOPOISOMERASE II beta.