Modulation of u-PA, MMPs and their inhibitors by a novel nutrient mixture in human glioblastoma cell lines

Human glioblastoma (LN-18, T-98G and A-172) cell lines (ATCC) were cultured in their respective media and treated at confluence with NM at 0, 50, 100, 250, 500 and 1000 µg/ml. Analysis of uPA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. Glioblastoma cell lines LN-18 and T-98G expressed uPA, which was inhibited by NM in a dose-dependent manner. However, no bands corresponding to uPA were detected for cell line A-172. On gelatinase zymography, all three cell lines showed bands corresponding to MMP-2 and LN-18 and T-98G showed PMA (100 ng/ml) -induced MMP-9. NM inhibited their expression in a dose-dependent manner. Activity of TIMP-2 was up regulated by NM in all glioma cell lines in a dose–dependent manner. Analysis revealed a positive correlation between uPA and MMP-2 and a negative correlation between uPA /MMPs and TIMP-2. These findings suggest the therapeutic potential of NM in treatment of gliomas.

Key words: 
glioblastoma LN-18, T-98G and A-172, uPA, MMP-2 and MMP-9, TIMP-2, PMA, nutrient mixture

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http://www.spandidos-publications.com/10.3892/ijo.2014.2465