Micro-array chip analysis of carbonyl-metabolising enzymes in normal, immortalised and malignant human oral keratinocytes.
Enzymes involved in various protective and metabolic processes of carbonyl compounds were analysed utilising a micro-array method in a three-stage in vitro model for oral carcinogenesis involving cultured normal, immortalised and malignant human oral keratinocytes. A complete transcript profiling of identified carbonyl-metabolising enzymes belonging to the ADH, ALDH, SDR and AKR families is presented. Expression of 17 transcripts was detected in normal, 14 in immortalized and 19 in malignant keratinocytes of a total of 12,500 genes spotted on the micro-array chip. For the detected transcripts, about half were changed by cell transformation, and for the various enzyme families, differences in expression patterns were observed. The detected AKR transcripts displayed a conserved pattern of expression, indicating a requirement for the keratinocyte phenotype, while most of the detected SDRs displayed changed expression at the various stages of malignancy. The importance of multiple experiments in using a microarray technique for reliable results is underlined and, finally, the strength of the method in detecting co-expressed enzymes in metabolic pathways is exemplified by the detection of the formaldehyde-scavenging pathway enzymes and the polyol pathway enzymes.
Hedberg
J J
JJ
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Grafström
R C
RC
Vondracek
M
M
Sarang
Z
Z
Wärngård
L
L
Höög
J O
JO
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D000818:Animals; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D009061:Mouth Mucosa / Q000166:cytology* / Q000201:enzymology; D009062:Mouth Neoplasms / Q000201:enzymology* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism
D016895:Culture Media, Serum-Free; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000426:Alcohol Dehydrogenase; D000449:Aldehyde Reductase; D000444:Aldehyde Dehydrogenase