title: Down-regulation of urokinase plasminogen activator and matrix metalloproteinases             and up-regulation of their inhibitors by a novel nutrient mixture in human prostate             cancer cell lines PC-3 and DU-145.
abstract: Strong clinical and experimental evidence shows that elevated levels of             urokinase plasminogen activators (u-PA) and matrix metalloproteinases (MMPs) are             associated with prostate cancer progression, metastasis and shortened survival             in patients. MMP activities are regulated by specific tissue inhibitors of metalloproteinases             (TIMPs). A nutrient mixture (NM) containing lysine, proline, ascorbic acid and             green tea extract showed anticancer activity against a number of cancer cell lines.             Our main objective was to study the effect of NM on the activity of u-PA, MMPs             and their inhibitor TIMPs on human prostate cancer cell lines PC-3 and DU-145.             Human prostate cancer cell lines PC-3 and DU-145 (ATCC) were grown in MEM media             with 10% FBS and antibiotics in 24‑well tissue culture plates. At near confluence,             the cells were treated with NM at 0-1000 µg/ml in triplicate at each concentration.             Analysis of u-PA activity was carried out by fibrin zymography, MMPs by gelatinase             zymography and TIMPs by reverse zymography. Both PC-3 and DU-145 prostate cancer             cell lines demonstrated u-PA activity (subunits 1 and 2, corresponding to 35 and             33 kDa). Prostate cancer cell line PC-3 secretion of u-PA subunit 1 was decreased             by 65% at NM 500 µg/ml and subunit 2 by 100% at NM 50 µg/ml. Prostate cancer cell             line DU-145 secretion of u-PA subunit 1 was decreased by 97% at NM 500 µg/ml and             subunit 2 by 100% at NM 100 µg/ml. Untreated PC-3 showed two bands for MMP-2 and             MMP-9. NM inhibited their expression in a dose-dependent manner. The activity             of MMP-2 and MMP-9 was significantly inhibited at 250 µg/ml with total inhibition             at 500 µg/ml. DU-145 cells did not exhibit MMP activity. Activity of TIMPs was             up-regulated in both prostate cancer cell lines in a dose-dependent manner. Minimum             activity was expressed at 50 µg/ml NM and maximum at 1000 µg/ml. Correlation analyses             revealed a positive correlation between u-PA and MMPs and a negative correlation             between u-PA/MMPs and TIMPs. These results suggest NM as a potential anticancer             agent since it targets invasive parameters of prostate cancer.
authors.affiliation: Dr Rath Research Institute, Cancer Division, Santa Clara, CA,                     USA.
authors.forename: M W
authors.initials: MW
authors.lastname: Roomi
authors.forename: T
authors.initials: T
authors.lastname: Kalinovsky
authors.forename: M
authors.initials: M
authors.lastname: Rath
authors.forename: A
authors.initials: A
authors.lastname: Niedzwiecki
mesh_terms: D045744:Cell Line, Tumor; D003470:Culture Media / Q000494:pharmacology*; D057075:Enzyme Assays; D015870:Gene Expression / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D018977:Micronutrients / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000378:metabolism*; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics / Q000378:metabolism*
chemical_list: D003470:Culture Media; D018977:Micronutrients; D019714:Tissue Inhibitor of Metalloproteinases; D014568:Urokinase-Type Plasminogen Activator; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9