Normal human fibroblasts enable melanoma cells to induce angiogenesis in type I collagen
Normal human fibroblasts enable melanoma cells to induce angiogenesis in type I collagen
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DOI：
10.1016/j.surg.2005.06.031
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发表时间：
2005-09-01
期刊：
影响因子：
3.8
通讯作者：
Velazquez, OC
中科院分区：
文献类型：
--
作者：
Goldstein, LJ;Chen, HY;Velazquez, OC
背景我们以前报道了成纤维细胞诱导人微血管内皮细胞（HMVECs）从单层分化为毛细血管样形态。我们现在测试的假设，即成纤维细胞调节血管生成的黑色素瘤细胞。我们测试了12个人黑色素瘤细胞系（2个径向生长期（RGP），3个垂直生长期（VGP）和7个转移性（MM））诱导HMVEC侵入/迁移到胶原蛋白中并形成毛细血管样网络的能力。HMVEC单层覆盖有单独的黑素瘤细胞（M）、单独的成纤维细胞（F）或2种细胞的1：1混合物（M + F）包埋的三维胶原蛋白。5天后，除去凝胶，固定，并通过冯维勒布兰德因子（nu MF）免疫荧光定量HMVEC网络。可溶性因子对HMVEC侵袭/迁移到胶原中的影响通过使用覆盖在HMVEC单层上的无细胞3-D胶原来评估，用源自M、F或M + F单层的条件培养基（CM）培养。通过使用RayBio细胞因子抗体阵列（RayBiotech，Norcross，Ga）鉴定参与观察到的侵袭/迁移的血管生成生长因子。仅当与成纤维细胞组合时观察到黑素瘤支持的血管生成中的细胞系特异性变异性（方差分析[ANOVA]，P <0.05）。01）。黑色素瘤加成纤维细胞一致地导致比单独的黑色素瘤显著更高的血管生成反应（P <0.05）。一个垂直生长期和一个转移性黑色素瘤系，虽然单独的血管生成弱，但当与成纤维细胞组合时，诱导的血管生成显著高于成纤维细胞或单独的黑色素瘤（P <0.05）。来自M或M + F的CM比单独来自成纤维细胞的CM诱导显著更少的HMVEC侵入/迁移到胶原中。白细胞介素8、单核细胞趋化蛋白-1和金属蛋白酶组织抑制剂-2在M + F培养物中的表达与单独的两种细胞类型相比显著升高。据我们所知，这是第一份报告表明，黑色素瘤支持的胶原血管生成更显着的影响，正常皮肤来源的成纤维细胞比黑色素瘤细胞类型的内在生物学。白细胞介素8，单核细胞趋化蛋白-1，和金属蛋白酶组织抑制剂-2牵连作为潜在的旁分泌因子调节这种观察到的效果。
Background. We previously reported that fibroblasts induce human microvascular endothelial cells (HMVECs) to differentiate from monolayer to capillarylike morphology. We now test the hypothesis that fibroblasts modulate angiogenesis in melanoma cells.Methods. We tested 12 human melanoma lines (2 radial growth phase (RGP), 3 vertical growth phase (VGP), and 7 metastatic (MM)) for ability to induce HMVECs to invade/migrate into collagen and form capillarylike networks. HMVEC monolayers were overlaid with 3-dimensional collagen gets embedded with melanoma cells alone (M), fibroblasts alone (F), or a 1:1 mixture of the 2 cells (M+F). After 5 days, gels were removed, fixed, and HMVEC networks were quantified by von Willebrand's factor (nu MF) immunofluorescence. The influence of soluble factors on HMVEC invasion migration into collagen was assessed with the use of acellulor 3-D collagen gets overlaid on HMVEC monolayers, cultured with conditioned media (CM) derived from nionolayers of M, F, or M+F Angiogenic growth factors involved in the observed invasionl/migration were identified with the use of a RayBio Cytokine Antibody Array (RayBiotech, Norcross, Ga).Results. Cell line-specific variability in melanoma-supported angiogenesis was observed only when in combination with fibroblasts (analysis of variance [ANOVA], P < . 01). Melanoma plus fibroblasts uniformly resulted in a signficantly higher angiogenic response than melanoma alone (P < .05). One vertical growth phase and one metastatic melanoma line, while weakly angiogenic alone, induced significantly higher angiogenesis than either fibroblast or melanoma alone (P < .05) when combined with fibroblasts. CM from M or M+F induced significantly less HMVEC invasion/migration into collagen than CM from fibroblasts alone. Interleukin 8, monocyte chemotactic protein-1, and tissue inhibitor Of metalloproteinase-2 were identified as signficantly elevated in the media derived from M+F cultures, compared with either cell type alone.Conclusion. To our knowledge, this is the first report demonstrating that melanoma-supported angiogenesis in collagen is more significantly influenced by normal skin-derived fibroblasts than by the intrinsic biology of the melanoma cell type. Interleukin 8, monocyte chemotactic protein-1, and tissue inhibitor of metalloproteinase-2 are implicated as potential paracrine factors regulating this observed effect.