title: Evaluation of cadmium-induced transcriptome alterations by three color cDNA labeling microarray analysis on a T-cell line.
abstract: Beside heavy metals, cadmium (Cd(2+)) is a ubiquitous toxic metal with a well established apoptotic and genotoxic effect, chronic exposure of which has been involved in a variety of pathological conditions. In the present study, we investigated by 1455 genes cDNA microarrays the toxic and apoptotic effect of Cd(2+), on the T-cell line CCRF-CEM, applying a three laser differential analysis, on the same microarray slide. The cells were cultured for 6 and 24 h in the absence (control) or presence of Cd(2+) (10 or 20 microM), RNAs were extracted and the produced cDNAs were labeled with rhodamine derivatives fluorescent dyes. A microarray slide was simultaneously hybridized by the labeled cDNAs and analyzed. We found that, in relation to control, treatment of the cells for 6 h with 10 and 20 microM Cd(2+), induces up-regulation in 20 and 34 genes, respectively. Treatment for 24 h with 10 and 20 microM Cd(2+) induces up-regulation in 22 and 84 genes, respectively. Twenty-eight genes were found down-regulated only after treatment for 24 h with Cd(2+) 10 microM. These data suggest that Cd(2+) produces a time- and dose-dependent molecular cascade, induces disturbances in different subcellular compartments, influencing thereafter the normal cellular functions, the differentiation process, the malignant transformation and the cell death.
authors.affiliation: University Research Institute for the Study and Treatment of Childhood Genetic and Malignant Diseases and Oncology Unit, First Department of Pediatrics, University of Athens, Aghia Sophia Childrens' Hospital, Greece. gthtsangaris@hotmail.com
authors.forename: George Th
authors.initials: GT
authors.lastname: Tsangaris
authors.forename: Athanassios
authors.initials: A
authors.lastname: Botsonis
authors.forename: Ioannis
authors.initials: I
authors.lastname: Politis
authors.forename: Fotini
authors.initials: F
authors.lastname: Tzortzatou-Stathopoulou
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D015152:Blotting, Northern; D002104:Cadmium / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics*; D005456:Fluorescent Dyes; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007202:Indicators and Reagents; D007834:Lasers; D009153:Mutagens / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D003470:Culture Media; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009153:Mutagens; D012333:RNA, Messenger; D002104:Cadmium