Microarray analysis of radiation response genes in primary human fibroblasts

Enikö Kis, Tünde Szatmári, Márton Keszei, Róbert Farkas, Olga Esik, Katalin Lumniczky, András Falus, Géza Sáfrány
International Journal of Radiation Oncology, Biology, Physics 2006 December 1, 66 (5): 1506-14

PURPOSE: To identify radiation-induced early transcriptional responses in primary human fibroblasts and understand cellular pathways leading to damage correction.

METHODS AND MATERIALS: Primary human fibroblast cell lines were irradiated with 2 Gy gamma-radiation and RNA isolated 2 h later. Radiation-induced transcriptional alterations were investigated with microarrays covering the entire human genome. Time- and dose dependent radiation responses were studied by quantitative real-time polymerase chain reaction (RT-PCR).

RESULTS: About 200 genes responded to ionizing radiation on the transcriptional level in primary human fibroblasts. The expression profile depended on individual genetic backgrounds. Thirty genes (28 up- and 2 down-regulated) responded to radiation in identical manner in all investigated cells. Twenty of these consensus radiation response genes were functionally categorized: most of them belong to the DNA damage response (GADD45A, BTG2, PCNA, IER5), regulation of cell cycle and cell proliferation (CDKN1A, PPM1D, SERTAD1, PLK2, PLK3, CYR61), programmed cell death (BBC3, TP53INP1) and signaling (SH2D2A, SLIC1, GDF15, THSD1) pathways. Four genes (SEL10, FDXR, CYP26B1, OR11A1) were annotated to other functional groups. Many of the consensus radiation response genes are regulated by, or regulate p53. Time- and dose-dependent expression profiles of selected consensus genes (CDKN1A, GADD45A, IER5, PLK3, CYR61) were investigated by quantitative RT-PCR. Transcriptional alterations depended on the applied dose, and on the time after irradiation.

CONCLUSIONS: The data presented here could help in the better understanding of early radiation responses and the development of biomarkers to identify radiation susceptible individuals.

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