Expression of different extracellular matrix components in human brain tumor and melanoma cells in respect to variant culture conditions
Expression of different extracellular matrix components in human brain tumor and melanoma cells in respect to variant culture conditions
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DOI：
10.1023/a:1006331416283
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发表时间：
1999-08-01
影响因子：
3.9
通讯作者：
Tonn, JC
中科院分区：
文献类型：
--
作者：
Bouterfa, H;Darlapp, AR;Tonn, JC
局部肿瘤侵犯周围脑组织是恶性胶质瘤的主要特征。这些过程很大程度上取决于肿瘤细胞与各种细胞外基质（ECM）成分的相互作用。由于关于 ECM 基因产物的一般表达和响应周围环境变化的表达的定量信息很少，因此设计了本研究。使用半定量 RT-PCR 测试了四种人类胶质母细胞瘤细胞系（U373MG、U138MG、U251MG、GaMG）以及四种人类黑色素瘤细胞系（MV3、BLM、530、IF6）表达不同人类 ECM 成分（纤连蛋白、核心蛋白聚糖、肌腱蛋白、I 型胶原蛋白、IV 型胶原蛋白、多功能蛋白聚糖）的 mRNA 的能力。此外，还分析了两种人髓母细胞瘤（MHH-Med 1、MHH-Med 4）和两种纤维肉瘤（HT1080、U2OS）细胞系。在含有 10% FCS 的 DMEM 培养基中生长的细胞表达大部分分析的蛋白质成分。当使用相同的培养基，但通过截止值 > 100 kD 的膜过滤来耗尽 ECM 蛋白时，大多数检查的细胞系中 ECM 蛋白的基础 mRNA 表达发生了变化。使用无血清条件，与其他培养基条件相比，大多数细胞系再次显示出编码不同 ECM 蛋白的 mRNA 表达模式的变化。比较来自一种肿瘤实体或不同肿瘤组的不同细胞系，不同肿瘤实体以及实体本身的 ECM 表达是异质的。迁移测定揭示了不同细胞系、ECM 成分和培养条件之间的异质反应，使得很难将 ECM 表达模式和迁移行为关联起来。我们的结果表明，所有检查的细胞系都能够在体外产生 ECM 蛋白。这表明肿瘤细胞可以在体外调节其微环境，在与迁移和侵袭相关的研究中必须考虑到这一点。
Local tumor invasion into the surrounding brain tissue is a major characteristic of malignant gliomas. These processes critically depend on the interaction of tumor cells with various extracellular matrix (ECM) components. Because only little quantitative information about expression of ECM gene products in general and expression in response to alterations of the surrounding environment is available, the present study was designed. Four human glioblastoma cell lines (U373MG, U138MG, U251MG, GaMG) as well as four human melanoma cell lines (MV3, BLM, 530, IF6) were tested with semiquantitative RT-PCR for their ability to express mRNA of different human ECM components (fibronectin, decorin, tenascin, collagen I, collagen IV, versican). In addition, two human medulloblastoma (MHH-Med 1, MHH-Med 4) and two fibrosarcoma (HT1080, U2OS) cell lines were analyzed. Cells which were grown in DMEM medium containing 10% FCS expressed most of the analyzed protein components. When the same medium, but depleted of ECM proteins by filtrating through a membrane with cut-off at > 100 kD was used, basal mRNA expression of the ECM proteins was changed in most of the examined cell lines. Using serum free conditions, most of the cell lines again showed a variation in the expression pattern of mRNA encoding for the different ECM proteins compared to the other medium conditions. Comparing different cell lines from one tumor entity or different tumor groups, ECM expression was heterogeneous with regard to the different tumor entities as well as within the entities themselves. Migration assays revealed heterogeneous responses between the different cell lines, ECM components and culture conditions, making it difficult to correlate ECM expression patterns and migratory behavior.Our results revealed that all examined cell lines are able to produce ECM proteins in vitro. This suggests that tumor cells can modulate their microenvironment in vitro which has to be taken into consideration for studies related to migration and invasion.