Effects of chlorpromazine with and without UV irradiation on gene expression of HepG2 cells.
Damage to DNA can trigger a variety of stress-related signals that alter the expression of genes associated with numerous biological pathways. In this study, we have used a cDNA microarray representing 1089 genes related to DNA damage and repair, cell cycle, transcription, metabolism and other toxicologically important cell functions to identify genes regulated in response to DNA damage in HepG2 cells induced by UV-activated chlorpromazine (CPZ). CPZ itself is not genotoxic but, upon UV irradiation with a non-genotoxic dose in the UVA range, it produces reactive free radical intermediates with DNA damaging properties. Genotoxicity in HepG2 cells was assessed concomitantly to gene expression profiling using the Comet assay. Kinetic studies were performed at a non-cytotoxic but clearly photogenotoxic concentration of CPZ (1.25 microg/ml) to characterize gene expression profiles at four different time points (3, 7, 15, 23 h) post short-term treatment. The results obtained from repeated experiments display a time-dependent expression pattern of up-regulated and repressed genes with distinct peaks in the number of differentially expressed genes at the 7 and 23 h time points. Most of the genes with altered expression belonged to the functional categories of cell cycle regulation and cell proliferation. A comparison with published expression profiles established in response to other genotoxic compounds showed low levels of concordance, which is most likely caused by the fact that extremely different testing conditions were used.
Frötschl
Roland
R
Federal Institute for Drugs and Medical Devices, Kurt-Georg-Kiesinger-Allee 3, 53175 Bonn, Germany. r.froetschl@bfarm.de
Weickardt
Sandra
S
Staszewski
Solveig
S
Kaufmann
Gabriele
G
Kasper
Peter
P
D000818:Animals; D045744:Cell Line, Tumor / Q000187:drug effects / Q000528:radiation effects; D002746:Chlorpromazine / Q000737:chemistry / Q000633:toxicity*; D020552:Comet Assay; D004249:DNA Damage; D053938:DNA Fragmentation; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000528:radiation effects*; D006801:Humans; D051379:Mice; D009153:Mutagens / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D025341:Principal Component Analysis; D013997:Time Factors; D014466:Ultraviolet Rays
D009153:Mutagens; D002746:Chlorpromazine