Microarray mRNA expression analysis of Fanconi anemia fibroblasts.
Fanconi anemia (FA) cells are generally hypersensitive to DNA cross-linking agents, implying that mutations in the different FANC genes cause a similar DNA repair defect(s). By using a customized cDNA microarray chip for DNA repair- and cell cycle-associated genes, we identified three genes, cathepsin B (CTSB), glutaredoxin (GLRX), and polo-like kinase 2 (PLK2), that were misregulated in untreated primary fibroblasts from three unrelated FA-D2 patients, compared to six controls. Quantitative real-time RT PCR was used to validate these results and to study possible molecular links between FA-D2 and other FA subtypes. GLRX was misregulated to opposite directions in a variety of different FA subtypes. Increased CTSB and decreased PLK2 expression was found in all or almost all of the analyzed complementation groups and, therefore, may be related to the defective FA pathway. Transcriptional upregulation of the CTSB proteinase appears to be a secondary phenomenon due to proliferation differences between FA and normal fibroblast cultures. In contrast, PLK2 is known to play a pivotal role in processes that are linked to FA defects and may contribute in multiple ways to the FA phenotype: PLK2 is a target gene for TP53, is likely to function as a tumor suppressor gene in hematologic neoplasia, and Plk2(-/-) mice are small because of defective embryonal development.
Galetzka
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D
Institute for Human Genetics, Johannes Gutenberg University, Mainz, Germany.
Weis
E
E
Rittner
G
G
Schindler
D
D
Haaf
T
T
D016022:Case-Control Studies; D002401:Cathepsin B / Q000235:genetics; D002453:Cell Cycle / Q000235:genetics; D003582:Cytogenetics; D004260:DNA Repair / Q000235:genetics; D005199:Fanconi Anemia / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D054477:Glutaredoxins / Q000235:genetics; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D054477:Glutaredoxins; D012333:RNA, Messenger; C496126:PLK2 protein, human; D017346:Protein Serine-Threonine Kinases; D002401:Cathepsin B