Unexpected effect of matrix metal loproteinase down-regulation on vascular intravasation and metastasis of human fibrosarcoma cells selected in vivo for high rates of dissemination
Unexpected effect of matrix metal loproteinase down-regulation on vascular intravasation and metastasis of human fibrosarcoma cells selected in vivo for high rates of dissemination
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DOI：
10.1158/0008-5472.can-05-2228
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发表时间：
2005-12-01
期刊：
影响因子：
11.2
通讯作者：
Quigley, JP
中科院分区：
文献类型：
--
作者：
Deryugina, EI;Zijlstra, A;Quigley, JP
将涉及将人HT-1080纤维肉瘤细胞移植到绒毛尿囊膜上的人肿瘤/鸡胚胎模型与定量实时Alu PCR结合使用，以在体内选择一对等基因细胞系（HT-hi/diss和HT-lo/diss），其从原发肿瘤扩散的能力显著不同（即，渗入绒毛尿囊膜血管系统并转移到肺部）。在免疫组织化学时程研究中，HT-hi/diss细胞依次从原发性肿瘤中逃逸，与血管接合，并最终在绒毛尿囊膜毛细血管内观察到，从而反映了早期的内渗事件。相比之下，HT-lo/diss细胞似乎仅限于其原发肿瘤。重要的是，在静脉内接种后，两种变体在宿主组织中以相似的效率停滞、外渗和增殖，这突出表明在原发性肿瘤外周观察到的早期事件可以解释它们的差异性传播。在这些事件的机制探测中，我们确定HT-hi/diss内渗对广泛的基质金属蛋白酶（AIMP）抑制剂敏感。为了分析单个MMP的可能作用，在HT-hi/diss细胞中用其相应的小干扰RNA分别下调膜结合MMP-14和分泌MMP-9。尽管MMP-14的有效下调，但HT-hi/diss细胞的浸润和转移均未受到显著影响。然而，MMP-9的显著下调伴随着血管内浸润和转移的惊人的3倍增加。结果强调了一种不断提高的认识，即靶向某些MMP可能导致增强的恶性肿瘤，在本文中以内渗水平为例，因为转移级联的这一步骤被解剖和量化。
The human tumor/chick embryo model involving grafting of human HT-1080 fibrosarcoma cells on the chorioallantoic membrane was used in conjunction with quantitative realtime Alu PCR to select in vivo a pair of isogenic cell lines (HT-hi/diss and HT-lo/diss), dramatically differing in their ability to disseminate from the primary tumor (i.e., intravasate into the chorioallantoic membrane vasculature and metastasize to the lungs). During an inummohistochemical time course study, HT-hi/diss cells were sequentially visualized having escaped from the primary tumors, engaged with the blood vessels, and eventually observed inside the chorioallantoic membrane capillaries, thus reflecting early intravasating events. In contrast, HT-lo/diss cells seemed restricted to their primary tumor. Importantly, after i.v. inoculation, both variants arrested, extravasated, and proliferated in host tissues with similar efficiencies, highlighting that the observed earlier events at the periphery of the primary tumor could account for their differential dissemination. In a mechanistic probing of these events, we determined that HT-hi/diss intravasation was sensitive to a broad-range matrix metalloproteinase (AIMP) inhibitor. To analyze the possible role of individual MMPs, membrane-bound MMP-14 and secreted MMP-9 were individually down-regulated in HT-hi/diss cells with their corresponding small interfering RNAs. Despite efficient down-regulation of MMP-14, neither intravasation nor metastasis of HT-hi/diss cells was affected significantly. However, a substantial down-regulation of MMP-9 was accompanied by a surprising 3-fold increase in intravasation and metastasis. The results emphasize a rising awareness that targeting certain MMPs might result in an enhanced malignancy, exemplified herein at the intravasation level as this step of the metastatic cascade is dissected and quantified.