Muir D, Johnson J, Rojiani M, Inglis B A, Rojiani A, Maria B L
Department of Pediatrics, University of Florida College of Medicine, Gainesville 32610, USA.
J Neurooncol. 1996 Dec;30(3):199-211. doi: 10.1007/BF00177271.
Cell motility within central nervous system (CNS) neuropil may be largely restricted yet infiltration by glioma cells is commonly observed. Glioma cells remodel nervous tissue and may assemble extracellular matrix in order to migrate. We examined the rat C6 glioma cell line for laminin expression and response in vitro and following engraftment into rat spinal cord. C6 cell cultures expressed laminin-2. C6 cells attached equally well to substrates of purified laminin-1 and laminin-2 and laminin-2-enriched C6 conditioned medium. In contrast, C6 cell migration was substantially greater on laminin-2 and C6-derived substrata than on laminin-1. Glioma cell attachment to laminin-1 and -2 was largely inhibited by antibody to the laminin receptor LBP110 and by an IKVAV peptide but not by YIGSR or control peptides. IKVAV peptide and anti-LBP110 antibodies also inhibited glioma cell invasion through synthetic basement membrane. Anti-beta 1 integrin antibody selectively inhibited cell migration and invasion on laminin-2 substrata without affecting percent cell attachment. These findings suggest C6 cell migration and invasion are promoted by autocrine release of laminin-2 and involve LPB110 and beta 1 integrin laminin receptors. A possible role for laminin-2 in CNS infiltration in vivo was examined following glioma engraftment into rat spinal cord. Engrafted C6 tumors share many histologic features of invasive human glioma. Engrafted glioma cells expressed laminin, LBP110 and beta 1 integrin antigens, indicating the molecular mechanisms of C6 motility observed in culture may contribute to glioma invasion in vivo. NMR and corroborative immunocytochemistry provided precise means to monitor tumor progression following glioma engraftment into rat spinal cord. Advantages of this glioma model are discussed regarding the assessment of anti-adhesive therapies in vivo.
中枢神经系统（CNS）神经纤维网内的细胞运动可能在很大程度上受到限制，但胶质瘤细胞的浸润却很常见。胶质瘤细胞重塑神经组织，并可能组装细胞外基质以便迁移。我们检测了大鼠C6胶质瘤细胞系在体外以及植入大鼠脊髓后的层粘连蛋白表达及反应。C6细胞培养物表达层粘连蛋白-2。C6细胞在纯化的层粘连蛋白-1、层粘连蛋白-2以及富含层粘连蛋白-2的C6条件培养基底物上的附着情况相同。相比之下，C6细胞在层粘连蛋白-2和C6来源的底物上的迁移明显大于在层粘连蛋白-1上的迁移。胶质瘤细胞与层粘连蛋白-1和-2的附着在很大程度上受到层粘连蛋白受体LBP110抗体和IKVAV肽的抑制，但不受YIGSR或对照肽的抑制。IKVAV肽和抗LBP110抗体也抑制胶质瘤细胞通过合成基底膜的侵袭。抗β1整合素抗体选择性抑制细胞在层粘连蛋白-2底物上的迁移和侵袭，而不影响细胞附着百分比。这些发现表明，层粘连蛋白-2的自分泌释放促进了C6细胞的迁移和侵袭，并且涉及LPB110和β1整合素层粘连蛋白受体。在将胶质瘤植入大鼠脊髓后，研究了层粘连蛋白-2在体内中枢神经系统浸润中的可能作用。植入的C6肿瘤具有侵袭性人类胶质瘤的许多组织学特征。植入的胶质瘤细胞表达层粘连蛋白、LBP110和β1整合素抗原，表明在培养中观察到的C6运动的分子机制可能有助于胶质瘤在体内的侵袭。核磁共振和确证性免疫细胞化学提供了精确的方法来监测胶质瘤植入大鼠脊髓后的肿瘤进展。讨论了该胶质瘤模型在体内评估抗粘连疗法方面的优势。
