Evaluation of cadmium-induced transcriptome alterations by three color cDNA labeling microarray analysis on a T-cell line.
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.
Tsangaris
George Th
GT
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
Botsonis
Athanassios
A
Politis
Ioannis
I
Tzortzatou-Stathopoulou
Fotini
F
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
D003470:Culture Media; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009153:Mutagens; D012333:RNA, Messenger; D002104:Cadmium