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

Our main objective was to study the effect of a nutrient mixture (NM) on activity of u-PA, MMPs and TIMPs on human lung and malignant mesothelioma (MM) cell lines. Human lung cancer (A-549 and Calu-3) and malignant mesothelioma (MSTO-211H) cell lines were cultured in their respective media and treated at confluence with NM at 0, 50, 100, 250, 500 and 1000 µg/ml. Analysis of u-PA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. Both lung cancer cell lines expressed u-PA, which was inhibited by NM in a dose-dependent manner. However, no bands corresponding to u-PA were detected for MM MSTO-211H cell line. On gelatinase zymography, A-549 showed one band corresponding to MMP-2, and induction of MMP-9 with PMA (100 ng/ml) treatment. MSTO-211H showed two bands, an intense band corresponding to MMP-2 and a faint band corresponding to MMP-9; MMP-9 was enhanced significantly with PMA treatment. NM inhibited their expression in both cell lines in a dose-dependent manner.  Calu-3 showed no MMP-2 or MMP-9 expression. Activity of TIMPs was up regulated by NM in all cancer cell lines in a dose–dependent manner. Analysis revealed a positive correlation between u-PA and MMPs and a negative correlation between u-PA /MMPs and TIMPs. These findings suggest the therapeutic potential of NM in treatment of lung and mesothelioma cancers.

Running Title: 
Nutrients modulate lung cancer cell u-PA, MMPs and TIMPs activity

Key words: 
lung cancer A-549 and Calu-3, malignant mesothelioma MSTO-211H, MMP-2 and MMP-9, nutrient mixture, TIMPs, u-PA