Hagemann Carsten, Anacker Jelena, Haas Stefanie, Riesner Daniela, Schömig Beate, Ernestus Ralf-Ingo, Vince Giles H
University of Würzburg, Department of Neurosurgery, Tumorbiology Laboratory, Würzburg, Germany.
BMC Res Notes. 2010 Nov 10;3:293. doi: 10.1186/1756-0500-3-293.
Glioblastomas (GBM), the most frequent malignant brain tumors in adults, are characterized by an aggressive local growth pattern and highly invasive tumor cells. This invasion is facilitated by expression of matrix metalloproteinases (MMPs), a family of zinc-dependent endopeptidases. They mediate the degradation of protein components of the extracellular matrix. Twenty-three family members are known. Elevated levels of several of them have been reported in GBM. GBM cell-lines are used for in vitro studies of cell migration and invasion. Therefore, it is essential to know their MMP expression patterns. Only limited data for some of the cell-lines are published, yet. To fill the gaps in our knowledge would help to choose suitable model systems for analysis of regulation and function of MMPs during GBM tumorigenesis, cell migration and invasion.
We analysed MMP-1, -8, -9, -10, -11, -13, -17, -19, -20, -21, -23, -24, -26, -27, and MMP-28 expression in seven GBM cell-lines (SNB-19, GaMG, U251, U87, U373, U343, U138) and in four primary cell cultures by semiquantitative RT-PCR, followed changes in the MMP expression pattern with increasing passages of cell culture and examined the influence of TNF-α and TGF-β1 stimulation on the expression of selected MMPs in U251 and U373 cells.MMP-13, -17, -19 and -24 were expressed by all analyzed cell-lines, whereas MMP-20 and MMP-21 were not expressed by any of them. The other MMPs showed variable expression, which was dependent on passage number. Primary cells displayed a similar MMP-expression pattern as the cell-lines. In U251 and U373 cells expression of MMP-9 and MMP-19 was stimulated by TNF-α. MMP-1 mRNA expression was significantly increased in U373 cells, but not in U251 cells by this cytokine. Whereas TGF-β1 had no impact on MMP expression in U251 cells, it significantly induced MMP-11 and MMP-24 expression in U373 cells.
Literature-data and our own results suggest that the expression pattern of MMPs is highly variable, dependent on the cell-line and the cell-culture conditions used and that also regulation of MMP expression by cytokines is cell-line dependent. This is of high impact for the transfer of cell-culture experiments to clinical implementation.
胶质母细胞瘤（GBM）是成人中最常见的恶性脑肿瘤，其特征为侵袭性的局部生长模式和具有高度侵袭性的肿瘤细胞。基质金属蛋白酶（MMPs）家族成员是一类锌依赖性内肽酶，其表达促进了这种侵袭。它们介导细胞外基质蛋白质成分的降解。已知该家族有23个成员。在GBM中已报道其中几种的水平升高。GBM细胞系用于细胞迁移和侵袭的体外研究。因此，了解它们的MMP表达模式至关重要。然而，目前仅公布了部分细胞系的有限数据。填补我们知识上的空白将有助于选择合适的模型系统，以分析GBM肿瘤发生、细胞迁移和侵袭过程中MMP的调节和功能。
我们通过半定量RT-PCR分析了7种GBM细胞系（SNB-19、GaMG、U251、U87、U373、U343、U138）和4种原代细胞培养物中MMP-1、-8、-9、-10、-11、-13、-17、-19、-20、-21、-23、-24、-26、-27和MMP-28的表达，跟踪细胞培养传代次数增加时MMP表达模式的变化，并检测了TNF-α和TGF-β1刺激对U251和U373细胞中所选MMP表达的影响。所有分析的细胞系均表达MMP-13、-17、-19和-24，而MMP-20和MMP-21均未在任何细胞系中表达。其他MMP表现出可变表达，这取决于传代次数。原代细胞显示出与细胞系相似的MMP表达模式。在U251和U373细胞中，TNF-α刺激了MMP-9和MMP-19的表达。该细胞因子使U373细胞中MMP-1 mRNA表达显著增加，但未使U251细胞中MMP-1 mRNA表达增加。而TGF-β1对U251细胞中的MMP表达没有影响，但它显著诱导了U373细胞中MMP-11和MMP-24的表达。
文献数据和我们自己的结果表明，MMP的表达模式高度可变，取决于所使用的细胞系和细胞培养条件，并且细胞因子对MMP表达的调节也具有细胞系依赖性。这对于将细胞培养实验转化为临床应用具有重要影响。
