Angiogenesis and tumor growth inhibition by a matrix metalloproteinase inhibitor targeting radiation-induced invasion
Angiogenesis and tumor growth inhibition by a matrix metalloproteinase inhibitor targeting radiation-induced invasion
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DOI：
10.1158/1535-7163.mct-05-0179
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发表时间：
2005-11-01
影响因子：
5.7
通讯作者：
Deutsch, E
中科院分区：
文献类型：
--
作者：
Kaliski, A;Maggiorella, L;Deutsch, E
在这项研究中，我们评估了电离辐射和基质金属蛋白酶（MMP）抑制剂之间的相互作用。使用Matrigel侵袭测定，我们表明电离辐射诱导培养的B16黑素瘤细胞的侵袭表型的剂量依赖性增加，并且来自这些经辐射的B16细胞的条件培养基促进人微血管内皮细胞（HMEC）的侵袭性。为了确定辐射诱导的侵袭性表型变化是否可能是由于MMP激活的变化引起的，我们使用体外黑色素瘤模型和小鼠皮下注射测试了MMP抑制剂Metastat调节电离辐射诱导的侵袭性表型的能力。肿瘤模型在这些研究中，Metastat抑制了培养的B16细胞中电离辐射诱导的侵袭表型，并类似地抑制了由来自辐射B16细胞的条件培养基诱导的HMEC侵袭的增加。相反，电离辐射增加B16 MMP-2的活性和照射B16诱导HMEC MMP-2活性的条件培养基。为了进一步研究电离辐射和MMP激活之间的相互作用，我们研究了电离辐射对MMP系统下游效应子的影响。我们发现，电离辐射诱导血管内皮生长因子（VEGF）分泌的B16黑色素瘤细胞，这种分泌被Metastat抑制。类似地，来自照射的B16的条件培养基也能够增加HMEC中的VEGF分泌。此外，电离辐射诱导的黑色素瘤细胞侵袭性被抗VEGF单克隆抗体部分抑制。在体内，电离辐射加上伴随的Metastat产生了黑色素瘤s.c.的最大生长抑制。肿瘤和这种效果与抑制血管生成，如多普勒超声和血小板/内皮细胞粘附分子-1染色测量。最后，电离辐射调制MMP-2，VEGF和VEGF受体表达在这些肿瘤样本中使用免疫组化。综上所述，这些结果表明，存在电离辐射诱导的肿瘤存活途径和可能的旁分泌电离辐射诱导的刺激途径，该途径从肿瘤细胞向上皮床发出，当同时给予Metastat时，该途径被阻碍。该模型可以提供MMP抑制剂与电离辐射结合以靶向辐射诱导的侵袭和血管生成的抗肿瘤功效的体内证据。
In this study, we have evaluated the interactions between ionizing radiation and a matrix metalloproteinase (MMP) inhibitor. Using Matrigel invasion assays, we show that ionizing radiation induced a dose-dependent increase in the invasive phenotype of cultured B16 melanoma cells and that conditioned medium from these irradiated B16 cells promoted enclothelial cell [human microvascular enclothelial cells (HMEC)l invasiveness. To determine whether the radiation-induced changes in invasive phenotype could be due to changes in MMP activation, we have tested the ability of the MMP inhibitor Metastat to modulate the ionizing radiation-induced invasive phenotype using both an in vitro melanoma model and a mouse s.c. tumor model. In these studies, Metastat inhibited the ionizing radiation-induced invasive phenotype in cultured B16 cells and similarly inhibited the increase in HMEC invasion induced by conditioned medium from irradiated B16 cells. Conversely, ionizing radiation increased B16 MMP-2 activity and the conditioned medium from irradiated B16 induced HMEC MMP-2 activity. To further investigate the interaction between ionizing radiation and MMP activation, we then studied the effects of ionizing radiation on downstream effectors of the MMP system. We found that ionizing radiation induced vascular endothelial growth factor (VEGF) secretion by B16 melanoma cells and that this secretion was inhibited by Metastat. Similarly, conditioned medium from irradiated B16 was also able to increase VEGF secretion in HMECs. Moreover, ionizing radiation -induced melanoma cell invasiveness was partially inhibited by an anti-VEGF monoclonal antibody. In vivo, ionizing radiation plus concomitant Metastat yielded the greatest growth inhibition of melanoma s.c. tumors and this effect correlated with inhibition of angiogenesis as measured by both Doppler ultrasonography and platelet/endothelial cell adhesion molecule-1 staining. Finally, ionizing radiation modulated MMP-2, VEGF, and VEGF receptor expression in these tumor samples using immunohistochemistry. Taken together, these results suggest that there is an ionizing radiationinduced tumor survival pathway and a possible paracrine ionizing radiation -induced stimulatory pathway emanating from tumor cells toward the enclothelial bed that is impeded when Metastat is given simultaneously. This model could provide in vivo evidence of the antitumor efficacy of combining a MMP inhibitor with ionizing radiation to target radiation-induced invasion and angiogenesis.