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Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
Published on: December 20, 2014
Diethyl sulfate induced Cdk2-dependent centrosome amplification in CHL cells
Entan Hu1, Juanling Fu, Peng Zhao
1Department of Toxicology, Peking University Health Science Center, Beijing 100191, PR China.
Abstract:
The function of centrosome that serves as the main microtubule organizing center is essential to ensure the genomic integrity during the cell division cycle. Centrosome abnormalities are frequently observed in many tumors and cells exposed to genotoxic agents. Here, we investigated the centrosome abnormalities induced by diethyl sulfate (DES) in Chinese hamster lung (CHL) fibroblasts and the underlying molecular mechanisms. Our results showed that DES exposure at 0.3, 1 and 3mM for 48 h caused centrosome amplification in a dose dependent manner. This effect was associated with transient S and G2/M phase delay and up-regulating of Cdk2, Cyclin A expressions. Furthermore, inhibition of Cdk2 activities reversed the centrosome amplification induced by DES. These results reveal that centrosome is one of the key subcellular targets of DES. Centrosome abnormalities might be important mechanisms behind the aneuploidy induction and carcinogenicity of DES.
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