Effect of Src kinase inhibition on metastasis and tumor angiogenesis in human pancreatic cancer
Effect of Src kinase inhibition on metastasis and tumor angiogenesis in human pancreatic cancer
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DOI：
10.1007/s10456-007-9071-3
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发表时间：
2007-01-01
期刊：
影响因子：
9.8
通讯作者：
Bruns, Christiane J.
中科院分区：
文献类型：
--
作者：
Ischenko, Ivan;Guba, Markus;Bruns, Christiane J.
肿瘤血管生成是一个需要内皮细胞迁移、增殖和分化的过程。我们假设，由于抑制Src活性而导致的胰腺肿瘤生长减少与Src激酶无法触发此类信号传导过程的网络相关，最终导致内皮细胞死亡和血管生成受限的肿瘤休眠。Src激酶抑制剂AZM 475271在裸鼠原位移植L3.6pl胰腺癌细胞中的治疗效果被测试。在AZM 475271治疗后未检测到肝转移和腹膜癌，并且观察到对平均胰腺肿瘤负荷的显著影响，这反过来与细胞增殖减少和凋亡内皮细胞增加相关。AZM 475271在体外Boyden Chamber细胞迁移试验中显示出显著抑制人脐静脉内皮细胞的迁移。在大鼠主动脉环试验中，我们也可以证明用类似剂量的Src激酶抑制剂治疗后对内皮细胞迁移和发芽的抑制。AZM 475271的最具决定性的抗血管生成活性在体内（小鼠角膜微囊试验）得到证实，显示A-ZM 475271全身给药后，碱性成纤维细胞生长因子诱导的新血管形成受到显著抑制。此外，我们可以显示用TACS MTT细胞活力测定试剂盒测定的HUVEC增殖降低。Src激酶的阻断显著降低了L3.6pl培养基中VEGF的水平，在来自HUVECs的细胞培养上清液中也发现了该效果。AZM 475271对Src激酶的抑制也显示了对VEGF和EGF受体的存活信号传导的阻止。用AZM 475271处理导致FAK酪氨酸磷酸化的VEGF依赖性抑制。还使用碘化丙啶染色通过流式细胞术检测HUVEC的细胞周期分析。Src激酶抑制剂以剂量依赖性方式促进HUVEC凋亡。综上所述，我们的研究结果表明，Src激酶抑制剂AZM 475271，除了其对肿瘤细胞的影响，抑制肿瘤的生长和转移，在体外和体内也可能通过抗血管生成机制。
Tumor angiogenesis is a process that requires migration, proliferation, and differentiation of endothelial cells. We hypothesized that decrease in pancreatic tumor growth due to inhibition of Src activity is associated with the inability of Src kinase to trigger a network of such signaling processes, which finally leads to endothelial cell death and angiogenesis-restricted tumor dormancy. The therapeutic efficacy of Src kinase inhibitor AZM475271 was tested in nude mice orthotopically xenografted with L3.6pl pancreatic carcinoma cells. No liver metastases and peritoneal carcinosis were detected and a significant effect on the average pancreatic tumor burden was observed following treatment with AZM475271, which in turn correlated with a decrease in cell proliferation and an increase in apoptotic endothelial cells. AZM475271 was shown to significantly inhibit migration of human umbilical vein endothelial cells in an in vitro Boyden Chamber cell migration assay. In a rat aortic ring assay we could demonstrate as well inhibition of endothelial cell migration and sprouting following therapy with Src kinase inhibitor at similar doses. The most conclusive anti-angiogenic activity of AZM475271 was demonstrated in vivo (mouse corneal micropocket assay) by showing a marked inhibition of basic fibroblast growth factor-induced neovascularization in response to systemic administration of A-ZM475271. Furthermore, we could show reduced proliferation of HUVECs determined with the TACS MTT Cell Viability Assay Kit. The blockade of Src kinase significantly reduced the level of VEGF in L3.6pl medium, the effect which was found also in the cell culture supernate from HUVECs. Inhibition of Src kinase by AZM475271 also showed prevention of survival signaling from VEGF and EGF receptors. Treatment with AZM475271 resulted in VEGF - dependent inhibition of tyrosine phosphorylation of FAK. HUVECs were also examined using propidium iodide staining for cell cycle analysis by FACS. Inhibition of Src kinase promoted HUVEC apoptosis in a dose-dependent manner. Taken together, our results suggest that the Src kinase inhibitor AZM475271, in addition to its effects on tumor cells, suppresses tumor growth and metastasis in vitro and in vivo potentially also by antiangiogenic mechanisms.