Human bladder cancer cells T-24 (ATCC) were grown in McCoy medium supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 mg/ml) in 24-well tissue culture plates. At near confluence, the cells were treated with the nutrient mixture (NM) dissolved in media and tested at 0, 10, 50, 100, 500, and 1000 mg/ml in triplicate at each dose. Cells were also treated with PMA 200 ng/ml to study enhanced MMP-9 activity. Cell proliferation was evaluated by MTT assay, MMP activity by gelatinase zymography, and invasion through Matrigel.
NM inhibited the T-24 cell secretion of MMP-2 and 9, with virtual total inhibition of MMP-2 at 500 mg/ml and MMP-9 at 100 µg/ml concentration. The nutrient mixture significantly reduced the invasion of human bladder cancer cells T-24 through Matrigel in a dose-dependent fashion, with 95% inhibition at 500 µg/ml and 100% at 1000 µg/ml NM (p<0.001).
Our results suggest that NM is an excellent candidate for therapeutic use in the treatment of bladder cancer, by inhibiting critical steps in cancer development and spread, such as MMP secretion and invasion.