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MAP kinases in proliferating human colon cancer Caco-2 cells

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Abstract

The mitogen-activated protein kinase (MAPK) cascade is one of the most ubiquitous signal transduction systems and is rapidly activated by various stimuli, such as cellular stress and death. The Caco-2 cell line is an in vitro model for colon cancer studies. We investigated the activation status of the ERK1/2, p38, JNK1/2, and ERK5 kinases and their respective upstream intracellular activators in Caco-2 cells induced to proliferate by 10% fetal bovine serum (FBS). The states of phosphorylation of the above MAPKs and their upstream kinases, MEK1/2, MKK3/6, MKK4, and MKK7, respectively, were studied by Western blot analysis. Phosphorylation was barely detectable before serum stimulation, and the stimulation of cell proliferation by the addition of FBS increased MEK1/2 and ERK1/2 phosphorylation 2 to 3 fold after 3 min. FBS stimulated p38 and MKK3/6 to the same extent within 2 min of treatment and JNK1/2 and its upstream kinases MKK4 and MKK7 5-fold (3 min). Addition of FBS also rapidly phosphorylated ERK5 (2 to 3.5-fold between 2 and 5 min) and the transcription factor CREB. Incubation of Caco-2 cells with FBS was followed by a rapid induction of c-Fos and c-Myc expression. Studies with ERK1/2 specific inhibitor PD98059, p38 MAPK inhibitor SB203580, or JNK inhibitor SP600125 showed that FBS regulates Caco-2 cell proliferation via the three MAPK pathways.

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Acknowledgements

This research was supported by grants from the Agencia Nacional de Promoción Cientifica y Tecnológica, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), and Universidad Nacional del Sur, Argentina.

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  1. Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, San Juan 670, 8000, Bahía Blanca, Argentina

    Natalia Buzzi, Andrea Colicheo, Ricardo Boland & Ana Russo de Boland

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  1. Natalia Buzzi
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  2. Andrea Colicheo
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  3. Ricardo Boland
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  4. Ana Russo de Boland
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Correspondence to Ana Russo de Boland.

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Buzzi, N., Colicheo, A., Boland, R. et al. MAP kinases in proliferating human colon cancer Caco-2 cells. Mol Cell Biochem 328, 201–208 (2009). https://doi.org/10.1007/s11010-009-0090-9

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