谷氨酸促脑胶质瘤细胞侵袭的机制研究

目的探讨谷氨酸促胶质瘤细胞侵袭的机制，检测MMP-2，MMP-9和uPAR的表达和激活。方法体外培养胶质瘤细胞株U251—MG，分别用50μM和100μM谷氨酸作用24后，Matrigel方法检测细胞侵袭；用含有100μM谷氨酸或同时含有谷氨酸受体阻断剂2pMMK-801的培养基作用U251-MG24小时，收集细胞用相对定量PCR检测MMP-2及MMP-9mRNA表达水平，收集上清后进行凝胶酶谱分析（ZymographyAssay）检测MMP-2、MMP-9的活性变化；检测谷氨酸刺激后uPAR的mRNA表达水平。结果谷氨酸浓度依赖的促进胶质瘤细胞株U251-MG侵袭；谷氨酸刺激没有改变U251-MG细胞中MMP-2和MMP-9表达水平，但使MMP-2活性增强，MMP-9活性没有变化；谷氨酸上调uPAR的表达。结论谷氨酸可能通过上调uPAR促进胶质瘤细胞发生MMP-2活性依赖的侵袭。

Study on the mechanism of invasion evoked by glutamine in cerebral glioma cells

Objective To investigate the mechanism of invasion evoked by glutamine in glioma and detect the expression and activation of MMP-2,MMP-9 and uPAR. Methods Glioma cell strain U251-MG was cultured in vitro.50 M and 100 M glutamine were respectively used to 24 hours. The cellular invasion was dected by Matrigel method.Contained glutamine receptor blocker 2 μM MK- 801 culture medium was used to U251-MG for 24 hours. After collecting cells,the expression levels of MMP-2 or MMP-9 mRNA were detected by relative quantitative PCR.Afler collecting supernate,the Zymography assay was performed and the active changes of MMP-2,MMP-9 were detected. Expression levels of mRNA of uPAR were detected after stimulating glutamine. Results Glutamine concentrationdependence promoted invasion of glioma cell strain U251-MG. Stimulation of glutamine didn＇t change expression levels of MMP-2 and MMP-9 in U251-MG cells. But the MMP-2 activity was enheanced and there was no change in the MMP-9 activity. Expression of uPAR was up-regulated by glutamine. Conclusion Glutamine may promote MMP-2 dependent invasion of glioma cells through up-regulatlon of uPAR.