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Endogenous Production of Nitric Oxide by Vascular Endothelial Growth Factor Down-Regulates Proliferation of Choriocarcinoma Cells

https://doi.org/10.1006/bbrc.2001.4682Get rights and content

Abstract

The trophoblast-like choriocarcinoma cell line BeWo expresses a receptor for vascular endothelial growth factor (VEGF) and proliferates in response to VEGF. Nitric oxide (NO) seems to play a key role in the VEGF-induced proliferation of endothelial cells but the NO mechanistic regulation of VEGF-stimulated trophoblast proliferation is presently unclear. We assessed the effect of exogenous VEGF on BeWo cell proliferation by [3H]thymidine incorporation. The VEGF-induced proliferation of BeWo cells was significantly increased by the NO synthase (NOS) inhibitor, Nω-nitro-l-arginine methyl ester (L-NAME), but was inhibited by the NO donor, sodium nitroprusside. Treatment of the cells with 10 ng/ml of VEGF increased not only eNOS expression but also NO production. The extracellular signal-regulated kinase (Erk) of the mitogen-activated protein kinase (MAPK) family was activated by VEGF as demonstrated by the phosphorylation of Erk in Western blots. The effects of VEGF on NO production and the expression of endothelial NOS (eNOS) were attenuated by treating BeWo cells with the selective inhibitor of MAPK kinase, PD98059. VEGF-stimulated proliferation of BeWo cells was inhibited by the tyrosine kinase inhibitor genistein but increased by PD98059. Other kinase inhibitors, including LY294002 (phosphoinositide 3-kinase inhibitor) and SB203580 (P38 MAPK inhibitor), had no effect on the proliferation of the cells and NO production. These results indicate that endogenous NO production down-regulates the VEGF-stimulated proliferation of BeWo cells and that the activation of Erk plays an important role in this mechanism.

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      The role of MAPKs in controlling proliferation of cell columns has not been tested, however, several growth factors controlling HTR-8/SVneo cell proliferation, i.e. TGF-α, amphiregulin or placental growth factor [66–68] likely act through ERK signalling. In addition, in choriocarcinoma cells lines angiotensin-II and vEGF modulate cell proliferation through ERK-dependent signalling [69–71]. BeWo cell proliferation is increased by leptin through ERK activation but this does not involve p38 MAPK [72].

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    Abbreviations used are: Erk, extracellular signal-regulated kinase; MAPK, mitogen-activated protein kinase; KDR, kinase-insert domain containing receptor; Flt-1, fms-like tyrosine kinase; L-NAME, Nω-nitro-l-arginine methyl ester; NO, nitric oxide; eNOS, endothelial nitric oxide synthase; SNP, sodium nitroprusside; VEGF, vascular endothelial growth factor.

    1

    To whom correspondence should be addressed at Department of Biochemistry, Dong-A University College of Medicine, 3 Ga-1, Dongdaeshin-Dong, Seo-Ku, Pusan 602-103, South Korea. Fax: 82-51-241-6940. E-mail: [email protected].

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