- 期刊
Colorimetric cDNA Microarray for Investigation of Transcription Profiles of Cells Exposed to a Therapeutic Arsenic Compound
並列摘要
Inorganic arsenic, a human carcinogen, is also an effective therapeutic agent for acute promyelocytic leukemia. Arsenic exposure induces numerous deletenious effects such as oxidative damages, chromosome aberrations, cell cycle arrest, and apoptosis in a variety of human cells. Unfortunately, the mechanism(s) of how arsenic induces apoptosis remains unclear. However, these cellular responses are greatly dependent on changes in gene expression. A recently developed colorimetric cDNA microarray technique allows us to investigate the changes in gene expression profiles of cells exposed to toxic substances. In this study, cDNA microarray membranes with 568 human genes were used to examine the transcripition profiling changes in human fibroblasts treated with 5 μM sodium arsenite for 0 to 24 h. Preliminary results showed that mRNA levels of heat shock proteins 60and 70 were time dependently increased in HFW cells, indicating the reliability of our assay system. Furthermore, mRNA levels of several cell cyclerelated genes, such as cyclin B1, cyclin G1 and DNA topoisomerase 11 d, were also elevated at late stage (12 -24 h) of arsenite treatment, confirming that arsenite could disturb cell cycle progression. mRNA levels of transcription factors and transcription regulators such as heat shock transcription factor, Egr-1, ATF2, GADD153, Zfp36, and SW1/SNF were also time-dependently increased in arsenite-treated HFW cells., In addition, the mRNA levels of several p53-activated genes such as p21/Cip, MDM-2, cyclin B1, cyclin G1, and GADD45 were up regulated after arsenite treatment, indicating p53 was activated in arsenite-treated HFW cells. These present results indicated that numerous specific transcription factors and regulators controlling cellular stress responses and cell cycle progression will be activated in cells responsible for toxic substance exposure.
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