Hov Håkon, Holt Randi Utne, Rø Torstein Baade, Fagerli Unn-Merete, Hjorth-Hansen Henrik, Baykov Vadim, Christensen James G, Waage Anders, Sundan Anders, Børset Magne
Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
Clin Cancer Res. 2004 Oct 1;10(19):6686-94. doi: 10.1158/1078-0432.CCR-04-0874.
We wanted to examine the role of the hepatocyte growth factor (HGF) receptor c-Met in multiple myeloma by applying a novel selective small molecule tyrosine kinase inhibitor, PHA-665752, directed against the receptor.
Four biological sequels of HGF related to multiple myeloma were studied: (1) proliferation of myeloma cells, (2) secretion of interleukin-11 from osteogenic cells, (3) migration of myeloma cells, and (4) adhesion of myeloma cells to fibronectin. We also examined effects of the c-Met inhibitor on intracellular signaling pathways in myeloma cells.
PHA-665752 effectively blocked the biological responses to HGF in all assays, with 50% inhibition at 5 to 15 nmol/L concentration and complete inhibition at around 100 nmol/L. PHA-665752 inhibited phosphorylation of several tyrosine residues in c-Met (Tyr(1003), Tyr(1230/1234/1235), and Tyr(1349)), blocked HGF-mediated activation of Akt and p44/42 mitogen-activated protein kinase, and prevented the adaptor molecule Gab1 from complexing with c-Met. In the HGF-producing myeloma cell line ANBL-6, PHA-665752 revealed an autocrine HGF-c-Met-mediated growth loop. The inhibitor also blocked proliferation of purified primary myeloma cells, suggesting that autocrine HGF-c-Met-driven growth loops are important for progression of multiple myeloma.
Collectively, these findings support the role of c-Met and HGF in the proliferation, migration, and adhesion of myeloma cells and identify c-Met kinase as a therapeutic target for treatment of patients with multiple myeloma.
我们希望通过应用一种新型的针对肝细胞生长因子（HGF）受体的选择性小分子酪氨酸激酶抑制剂PHA-665752，来研究HGF受体c-Met在多发性骨髓瘤中的作用。
研究了与多发性骨髓瘤相关的HGF的四个生物学后续效应：（1）骨髓瘤细胞的增殖，（2）成骨细胞白细胞介素-11的分泌，（3）骨髓瘤细胞的迁移，以及（4）骨髓瘤细胞与纤连蛋白的黏附。我们还研究了c-Met抑制剂对骨髓瘤细胞内信号通路的影响。
PHA-665752在所有实验中均有效阻断了对HGF的生物学反应，在5至15 nmol/L浓度时抑制率达50%，在约100 nmol/L时完全抑制。PHA-665752抑制了c-Met中几个酪氨酸残基（Tyr(1003)、Tyr(1230/1234/1235)和Tyr(1349)）的磷酸化，阻断了HGF介导的Akt和p44/42丝裂原活化蛋白激酶的激活，并阻止衔接分子Gab1与c-Met结合。在产生HGF的骨髓瘤细胞系ANBL-6中，PHA-665752揭示了一种自分泌HGF-c-Met介导的生长环。该抑制剂还阻断了纯化的原代骨髓瘤细胞的增殖，表明自分泌HGF-c-Met驱动的生长环对多发性骨髓瘤的进展很重要。
总体而言，这些发现支持了c-Met和HGF在骨髓瘤细胞增殖、迁移和黏附中的作用，并确定c-Met激酶是治疗多发性骨髓瘤患者的一个治疗靶点。
