Identification of genes responsive to gamma radiation in rat hepatocytes and rat liver by cDNA array gene expression analysis.
The mechanisms underlying hepatocellular damage after irradiation are obscure. We identified genes induced by radiation in isolated rat hepatocytes in vitro by cDNA array gene expression analysis and then screened in vivo experiments with those same genes using real-time PCR and Western blotting. Hepatocytes were irradiated and cDNA array analyses were performed 6 h after irradiation. The mRNA of differentially expressed genes was quantitatively analyzed by real-time PCR. cDNA array analyses showed an up-regulation of 10 genes in hepatocytes 6 h after irradiation; this was confirmed by real-time PCR. In vivo, rat livers were irradiated selectively. Treated and sham-irradiated controls were killed humanely 1, 3, 6, 12, 24 and 48 h after irradiation. Liver RNA was analyzed by real-time PCR; expression of in vivo altered genes was also analyzed at the protein level by Western blotting. Up-regulation was confirmed for three of the in vitro altered genes (multidrug resistance protein, proteasome component C3, eukaryotic translation initiation factor 2). Histologically, livers from irradiated animals were characterized by steatosis of hepatocytes. Thus we identified genes that may be involved in liver steatosis after irradiation. The methods shown in this work should help to further clarify the consequences of radiation exposure in the liver.
Christiansen
Hans
H
Department of Radiotherapy, Goettingen University, 37075 Goettingen, Germany. hchrist@gwdg.de
Batusic
Danko
D
Saile
Bernhard
B
Hermann
Robert Michael
RM
Dudas
Josef
J
Rave-Frank
Margret
M
Hess
Clemens Friedrich
CF
Schmidberger
Heinz
H
Ramadori
Giuliano
G
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D005720:Gamma Rays; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000528:radiation effects*; D022781:Hepatocytes / Q000378:metabolism* / Q000528:radiation effects*; D008099:Liver / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects*; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D015852:Eukaryotic Initiation Factor-2; D012333:RNA, Messenger