Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, Massachusetts, USA.
Clin Cancer Res. 2011 Jul 1;17(13):4389-99. doi: 10.1158/1078-0432.CCR-10-2772. Epub 2011 Apr 26.
There is no standard of therapy for the treatment of Waldenström macroglobulinemia (WM), therefore there is a need for the development of new agents. Fibroblast growth factor receptor 3 (FGFR3) was shown to play a major role in several types in cancer. Dovitinib, an inhibitor of FGFR3, was effective in hematologic malignancies. In this study, we tested FGFR3 as a therapeutic target in WM and tested the effect of dovitinib on cell proliferation and apoptosis of WM cells in the context of BM microenvironment.
The expression of FGFR3 in WM cells was tested using immunofluorescence and flow cytometry. Cell signaling in response to stimulation with FGF3 and stromal cells, and its inhibition by dovitinib was performed using immunoblotting. Cell survival and cell proliferation were assessed by MTT and BrdU assays. Apoptosis was measured by detection of APO-2.7 and cleavage of caspase-3 using flow cytometry. Cell cycle was performed by PI staining of cells and flow cytometry. The combinatory effect of dovitinib with other drugs was analyzed using Calcusyn software. The effect of dovitinib was tested in vivo.
FGFR3 was overexpressed in WM cells and its activation induced cell proliferation. Inhibition of FGFR3 with dovitinib decreased cell survival, increased apoptosis, and induced cell cycle arrest. Inhibition of FGFR3 by dovitinib reduced the interaction of WM to bone marrow components, and reversed its proliferative effect. Dovitinib had an additive effect with other drugs. Moreover, dovitinib reduced WM tumor progression in vivo.
We report that FGFR3 is a novel therapeutic target in WM, and suggest dovitinib for future clinical trial the treatment of patients with WM.
目前针对 Waldenström 巨球蛋白血症（WM）的治疗尚无标准疗法，因此需要开发新的药物。成纤维细胞生长因子受体 3（FGFR3）在多种类型的癌症中发挥着重要作用。成纤维细胞生长因子受体 3 抑制剂多韦替尼在血液恶性肿瘤中具有疗效。在这项研究中，我们检测了 FGFR3 作为 WM 的治疗靶点，并检测了多韦替尼在骨髓微环境下对 WM 细胞增殖和凋亡的影响。
使用免疫荧光和流式细胞术检测 WM 细胞中 FGFR3 的表达。使用免疫印迹法检测 FGFR3 对 FGF3 和基质细胞刺激的信号转导及其对多韦替尼的抑制作用。通过 MTT 和 BrdU 测定评估细胞存活和增殖。通过流式细胞术检测 APO-2.7 的表达和 caspase-3 的裂解来测量细胞凋亡。通过细胞 PI 染色和流式细胞术检测细胞周期。使用 Calcusyn 软件分析多韦替尼与其他药物的联合效应。在体内检测多韦替尼的作用。
FGFR3 在 WM 细胞中过度表达，其激活可诱导细胞增殖。多韦替尼抑制 FGFR3 可降低细胞存活率，增加细胞凋亡，并诱导细胞周期停滞。多韦替尼抑制 FGFR3 可减少 WM 与骨髓成分的相互作用，并逆转其增殖作用。多韦替尼与其他药物具有相加作用。此外，多韦替尼可减少体内 WM 肿瘤的进展。
我们报告 FGFR3 是 WM 的一个新的治疗靶点，并建议将多韦替尼用于未来治疗 WM 患者的临床试验。
