Mapping of oxidative stress responses of human tumor cells following photodynamic therapy using hexaminolevulinate.
BACKGROUND
Photodynamic therapy (PDT) involves systemic or topical administration of a lesion-localizing photosensitizer or its precursor, followed by irradiation of visible light to cause singlet oxygen-induced damage to the affected tissue. A number of mechanisms seem to be involved in the protective responses to PDT, including activation of transcription factors, heat shock proteins, antioxidant enzymes and apoptotic pathways.


RESULTS
In this study, we address the effects of a destructive/lethal hexaminolevulinate (HAL) mediated PDT dose on the transcriptome by using transcriptional exon evidence oligo microarrays. Here, we confirm deviations in the steady state expression levels of previously identified early defence response genes and extend this to include unreported PDT inducible gene groups, most notably the metallothioneins and histones. HAL-PDT mediated stress also altered expression of genes encoded by mitochondrial DNA (mtDNA). Further, we report PDT stress induced alternative splicing. Specifically, the ATF3 alternative isoform (deltaZip2) was up-regulated, while the full-length variant was not changed by the treatment. Results were independently verified by two different technological microarray platforms. Good microarray, RT-PCR and Western immunoblotting correlation for selected genes support these findings.


CONCLUSIONS
Here, we report new insights into how destructive/lethal PDT alters the transcriptome not only at the transcriptional level but also at post-transcriptional level via alternative splicing.
Cekaite
Lina
L
Department of Tumor Biology, Rikshopitalet - Radiumhospitalet Medical Center, 0310 Oslo, Norway. linac@rr-research.no
Peng
Qian
Q
Reiner
Andrew
A
Shahzidi
Susan
S
Tveito
Siri
S
Furre
Ingegerd E
IE
Hovig
Eivind
E
D051700:Activating Transcription Factor 3 / Q000378:metabolism; D017398:Alternative Splicing / Q000187:drug effects; D000622:Aminolevulinic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004272:DNA, Mitochondrial / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005091:Exons; D005786:Gene Expression Regulation / Q000187:drug effects; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D015458:Leukemia, T-Cell / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D048670:MAP Kinase Kinase 4 / Q000235:genetics / Q000378:metabolism; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000187:drug effects*; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000494:pharmacology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
C490449:ATF3 protein, human; D051700:Activating Transcription Factor 3; D004272:DNA, Mitochondrial; D006657:Histones; D017319:Photosensitizing Agents; D020033:Protein Isoforms; D016271:Proto-Oncogene Proteins c-myc; D000622:Aminolevulinic Acid; D048670:MAP Kinase Kinase 4; D045726:Metalloproteases; C419924:5-aminolevulinic acid hexyl ester
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