Endogenous production of nitric oxide by vascular endothelial growth factor down-regulates proliferation of choriocarcinoma cells
Endogenous production of nitric oxide by vascular endothelial growth factor down-regulates proliferation of choriocarcinoma cells
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DOI：
10.1006/bbrc.2001.4682
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发表时间：
2001-04-13
影响因子：
3.1
通讯作者：
Kwak, JY
中科院分区：
文献类型：
--
作者：
Cha, MS;Lee, MJ;Kwak, JY
滋养层细胞样绒癌细胞系BeWo表达血管内皮生长因子受体，并在血管内皮生长因子的作用下增殖。一氧化氮(NO)似乎在血管内皮生长因子(VEGF)诱导的内皮细胞增殖中起关键作用，但NO对血管内皮生长因子刺激的滋养层细胞增殖的调节机制目前尚不清楚。通过[H-3]胸腺嘧啶核苷掺入法检测外源性血管内皮生长因子对BeWo细胞增殖的影响。一氧化氮合酶抑制剂L精氨酸甲酯可显著促进血管内皮生长因子诱导的BeWo细胞增殖，而NO供体硝普钠则抑制其增殖。10 ng/ml的血管内皮细胞生长因子不仅能增加eNOS的表达，还能增加NO的产生。丝裂原活化蛋白激酶(MAPK)家族中的细胞外信号调节激酶(ERK)被血管内皮生长因子(VEGF)激活，Western blots中ERK的磷酸化证明了这一点。选择性MAPK激酶抑制剂PD98059处理BeWo细胞后，可减弱血管内皮生长因子对细胞NO生成和内皮型一氧化氮合酶(ENOS)表达的影响，酪氨酸激酶抑制剂Genistein可抑制VEGF刺激的BeWo细胞增殖，而PD98059可促进其增殖，而磷酸肌醇3-激酶抑制剂LY294002和P38 MAPK抑制剂SB203580对细胞增殖和NO生成无明显影响。这些结果表明，内源性NO的产生下调了血管内皮生长因子刺激的BeWo细胞的增殖，ERK的激活在此机制中起重要作用。(C)2001年学术出版社。
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 [H-3]thymidine incorporation. The VEGF-induced proliferation of BeWo cells was significantly increased by the NO synthase (NOS) inhibitor, N-omega-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. (C) 2001 Academic Press.