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Molecular Characterization of STAT Signaling in Inflammation and Tumorigenesis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 512))

Summary

The Janus kinases (JAK) and signal transducer and activator of transcription (STAT) signaling are strongly activated in many tumors. STAT proteins are activated by phosphorylation at the tyrosine residue, then dimerize, translocate to the nucleus and bind DNA, initiating the transcription of target genes. Activation of JAK-STAT pathway is implicated in the regulation of cell growth, differentiation, survival and cross-talk between cancer and immune cells.

The activation of STATs depends on phosphorylation on a single tyrosine residue (e.g., Tyr705 in STAT3 and Tyr694 in STAT5) in the C-terminal domain. Commercially available antibodies discriminate between total and specifically phosphorylated (active) forms of different STATs, which allows to measure directly STATs activation in crude cell extracts. Nuclear translocation and transcriptional activity of STATs can be measured in transfected cells using STAT dependent promoter driving reporter luciferase gene. STAT signaling pathway and STAT-dependent gene expression in cells can be specifically modulated using oligodeoxynucleotide (ODN) STAT decoy which is a double-stranded fragment of DNA containing an overlapping ISRE/GAS binding site.

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Acknowledgments

This work was supported by Ministry of Science and Higher Education (PBZ-MIN 014/P05/2004).

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Adach, A., Ellert-Miklaszewska, A., Kaminska, B. (2009). Molecular Characterization of STAT Signaling in Inflammation and Tumorigenesis. In: Kozlov, S.V. (eds) Inflammation and Cancer. Methods in Molecular Biology™, vol 512. Humana Press. https://doi.org/10.1007/978-1-60327-530-9_14

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  • DOI: https://doi.org/10.1007/978-1-60327-530-9_14

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-529-3

  • Online ISBN: 978-1-60327-530-9

  • eBook Packages: Springer Protocols

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