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Repression of STAT3, STAT5A, and STAT5B expressions in chronic myelogenous leukemia cell line K–562 with unmodified or chemically modified siRNAs and induction of apoptosis

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Abstract

Signal transducers and activators of transcription (STAT) proteins are latent cytoplasmic transcription factors that affect several cellular processes including cell growth, proliferation, differentiation, and survival. Following phosphorylation, STATs are activated, and their upregulated expressions increase in malignancies with playing a role in the development of leukemia. In this study, transfection of K–562 cells with either unmodified or chemically modified anti-STAT3, -STAT5A, -STAT5B siRNAs for duration of 12 days, determining gene silencing at mRNA and protein levels, evaluating apoptosis rate, and detecting JAK/STAT pathway members’ gene expression profiles via array method were aimed. Quantitative RT-PCR and Western blot assays indicated that STAT expressions were downregulated both at mRNA and protein levels, and TUNEL assay showed that leukemic cell apoptosis was induced due to inhibition of STATs. Array analysis resulted with decreases in signal transducer, phosphorylation inducer, and oncogene expressions, whereas increased expressions in STAT inhibitor and apoptosis inducer genes were observed. These results point out that siRNA application could constitute a new and alternative curative method for supporting therapy of CML-diagnosed patients in the future.

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Acknowledgments

We would like to thank Dr. Vildan Bozok ÇETİNTAŞ, Phd (Ege University Medical School, of Medical Biology Department) for the great help in creating figures of the manuscript. This work was supported by The Scientific and Technological Research Council of Turkey, by the Turkish Society of Hematology, and Ege University Medical School Scientific Research Projects (TUBITAK 105S459, THD 200–02/08 and APAK 2010-TIP–004 respectively, to BK).

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The authors declare no potential conflict of interest.

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Authors and Affiliations

  1. Faculty of Medicine Department of Medical Biology, Ege University, 35100, Bornova, Izmir, Turkey

    Burçin Tezcanlı Kaymaz, Nur Selvi, Cumhur Gündüz, Çağdaş Aktan, Ayşegül Dalmızrak & Buket Kosova

  2. Faculty of Medicine Department of Internal Medicine–Hematology, Ege University, Izmir, Turkey

    Güray Saydam

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  1. Burçin Tezcanlı Kaymaz
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  2. Nur Selvi
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Correspondence to Burçin Tezcanlı Kaymaz.

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Kaymaz, B.T., Selvi, N., Gündüz, C. et al. Repression of STAT3, STAT5A, and STAT5B expressions in chronic myelogenous leukemia cell line K–562 with unmodified or chemically modified siRNAs and induction of apoptosis. Ann Hematol 92, 151–162 (2013). https://doi.org/10.1007/s00277-012-1575-2

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  • DOI: https://doi.org/10.1007/s00277-012-1575-2

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