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B-CLL

Flavopiridol downregulates the expression of both the inducible NO synthase and p27kip1 in malignant cells from B-cell chronic lymphocytic leukemia

Leukemia volume 17, pages 2435–2443 (2003)Cite this article

Abstract

Flavopiridol, an inhibitor of cyclin-dependent kinases and other protein kinases, induces in vitro apoptosis of malignant cells from B-cell chronic lymphocytic leukemia (B-CLL). Previously, we reported that nitric oxide (NO), produced by an inducible NO synthase (iNOS), spontaneously expressed by the B-CLL cells, contributed to their deficiency in apoptosis. In the present work, we show that ex vivo treatment of leukemic cells from B-CLL patients with flavopiridol results in the inhibition of iNOS expression, as determined by immunofluorescence and Western blotting, and in a marked inhibition of NO production measured in situ with a specific fluorescent probe (DAF-2 DA). These effects are accompanied by membrane, mitochondrial and nuclear events of apoptosis. Flavopiridol exposure also results in the stimulation of caspase 3 activity and in caspase-dependent cleavage of p27kip1, a negative regulator of the cell cycle, which is overexpressed in B-CLL. Thus, flavopiridol is capable of downregulating both iNOS and p27kip1 expression in B-CLL cells. Furthermore, flavopiridol-promoted apoptosis is partly reverted by an NO donor, suggesting that inhibition of the NO pathway could participate in the apoptotic effects of flavopiridol on the leukemic cells.

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Acknowledgements

We are very grateful to Mrs D Rouillard for her expert FACS analysis. We thank Dr JR Suarez (Aventis Pharmaceuticals) for his generous gift of flavopiridol. This work was supported by INSERM, and a grant from the Fondation contre la Leucémie.

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Authors and Affiliations

  1. U 365 INSERM, Institut Curie, Paris, France

    C Billard, C Kern & J-P Kolb

  2. E 355 INSERM, Hôpital Hôtel-Dieu, Paris, France

    R Tang & F Ajchenbaum-Cymbalista

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Billard, C., Kern, C., Tang, R. et al. Flavopiridol downregulates the expression of both the inducible NO synthase and p27kip1 in malignant cells from B-cell chronic lymphocytic leukemia. Leukemia 17, 2435–2443 (2003). https://doi.org/10.1038/sj.leu.2403139

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