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

Roomi,M.  Waheed; Kalinovsky,Tatiana; Niedzwiecki,Aleksandra; Rath,Matthias

doi:10.3892/ijo.2013.1934

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

Authors:
- M. Waheed Roomi
- Tatiana Kalinovsky
- Aleksandra Niedzwiecki
- Matthias Rath
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Affiliations: Dr Rath Research Institute, Santa Clara, CA 95050, USA
Published online on: May 9, 2013 https://doi.org/10.3892/ijo.2013.1934
Pages: 39-49
Copyright: © Roomi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Adult sarcomas are highly aggressive tumors that are characterized by high levels of matrix metalloproteinase (MMP)-2 and -9 secretions that degrade the ECM and basement membrane, allowing cancer cells to spread to distal organs. Proteases play a key role in tumor cell invasion and metastasis by digesting the basement membrane and ECM components. Strong clinical and experimental evidence demonstrates association of elevated levels of u-PA and MMPs with cancer progression, metastasis and shortened patient survival. MMP activities are regulated by specific tissue inhibitors of metalloproteinases (TIMPs). Our main objective was to study the effect of a nutrient mixture (NM) on the activity of u-PA, MMPs and TIMPs in various human adult sarcomas. Human fibrosarcoma (HT-1080), chondrosarcoma (SW-1353), liposarcoma (SW-872), synovial sarcoma (SW-982) and uterine leimyosarcoma (SK-UT-1) cell lines (ATCC) were cultured in their respective media and treated at confluence with NM at 0, 50, 100, 250, 500 and 1,000 µg/ml. Analysis of u-PA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. Fibrosarcoma, chondrosarcoma, liposarcoma and leiomyosarcoma 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 synovial sarcoma cells. On gelatinase zymography, fibrosarcoma, chondrosarcoma, liposarcoma and synovial sarcoma showed bands corresponding to MMP-2 and MMP-9 with enhancement of MMP-9 with PMA (100 ng/ml) treatment. Uterine leiomyosarcoma showed strong bands corresponding to inactive and active MMP-9 and a faint band corresponding to MMP-9 dimer induced with PMA treatment, but no MMP-2 band. NM inhibited their expression in a dose‑dependent manner. Activity of TIMPs was upregulated 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 adult sarcomas.

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1.	American Cancer Society: Adult soft tissue cancer. What are key statistics of soft tissue cancers? http://www.cancer.org/cancer/sarcoma-adultsofttissuecancer/detailedguide/sarcoma-adult-soft-tissue-cancer-key-statistics. Accessed January 21, 2013.
2.	American Cancer Society: Bone cancer. What are the key statistics about bone cancer? http://www.cancer.org/cancer/bonecancer/detailedguide/bone-cancer-key-statistics. Accessed January 21, 2013.
3.	Papagelopoulos PJ, Galanis E, Frassica FJ, Sim FH, Larson DR and Wold LE: Primary fibrosarcoma of bone. Outcome after primary surgical treatment. Clin Orthop Relat Res. 373:88–103. 2000.PubMed/NCBI
4.	Benassi MS, Gamberi G, Magagnoli G, Molendini L, Ragazzini P, Merli M, Chiesa F, Balladelli A, Manfrini M, Bertoni F, Mercri M and Picci P: Metalloproteinase expression and prognosis in soft tissue sarcomas. Ann Oncol. 12:75–80. 2001. View Article : Google Scholar : PubMed/NCBI
5.	Fidler IJ: Molecular biology of cancer: invasion and metastasis. Cancer Principles and Practice of Oncology. De Vita VT, Hellman S and Rosenberg SA: 5th edition. Lippincott-Raven; Philadelphia, PA: pp. 135–152. 1997
6.	Egeblad M and Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2:161–174. 2002. View Article : Google Scholar : PubMed/NCBI
7.	Folkman J: Role of angiogenesis in tumor growth and metastasis. Semin Oncol. 29(Suppl 16): 15–18. 2002. View Article : Google Scholar : PubMed/NCBI
8.	Chambers AF and Matrisian LM: Changing views on the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst. 89:1260–1270. 1997. View Article : Google Scholar : PubMed/NCBI
9.	Kleiner DL and Stetler-Stevenson WG: Matrix metalloproteinases and metastasis. Cancer Chemother Pharmacol. 43(Suppl): 42s–51s. 1999. View Article : Google Scholar
10.	Yurchenko PD and Schitny JC: Molecular architecture of basement membranes. FASEB J. 4:1577–1590. 1990.PubMed/NCBI
11.	Barsky SH, Siegel GP, Jannotta F and Liotta LA: Loss of basement membrane components by invasive tumors but not by their benign counterparts. Lab Investig. 49:140–147. 1983.PubMed/NCBI
12.	Liotta LA, Tryggvason K, Garbisa A, Hart I, Foltz CM and Shafie S: Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature. 284:67–68. 1980. View Article : Google Scholar : PubMed/NCBI
13.	Nelson AR, Fingleton B, Rothenberg ML and Matrisian LM: Matrix metalloproteinases: biologic activity and clinical implications. J Clin Oncol. 18:1135–1149. 2000.PubMed/NCBI
14.	Taubert H, Würl P, Greither T, Kappler M, Bache M, Lautenschläger C, Füssel S, Meye A, Eckert AW, Holzhausen HJ, Magdolen V and Kotzsch M: Co-detection of members of the urokinase plasminogen activator system in tumour tissue and serum correlates with a poor prognosis for soft-tissue sarcoma patients. Br J Cancer. 102:731–737. 2010. View Article : Google Scholar : PubMed/NCBI
15.	Stetler-Stevenson WG: The role of matrix metalloproteinases in tumor invasion, metastasis and angiogenesis. Surg Oncol Clin N Am. 10:383–392. 2001.PubMed/NCBI
16.	Stetler-Stevenson WG: Type IV collagenases in tumor invasion and metastasis. Cancer Metastasis Rev. 9:289–303. 1990. View Article : Google Scholar : PubMed/NCBI
17.	Benassi MS, Magagnoli G, Ponticelli F, Pazzaglia L, Zanella L, Gamberti G, Ragazzini P, Ferrari C, Mercuri M and Picci P: Tissue and serum loss of metalloproteinases inhibitors in high-grade soft tissue sarcomas. Histol Histopathol. 18:1035–1040. 2003.PubMed/NCBI
18.	Dano K, Andreasen PA, Grondahl-Hansen J, Kristensen P, Nielsen LS and Skriver L: Plasminogen activators, tissue degradation and cancer. Adv Cancer Res. 44:139–266. 1985. View Article : Google Scholar : PubMed/NCBI
19.	Duffy MJ, Duggan C, Mulcahy HE, McDermott EW and O'Higgins NJ: Urokinase plasminogen activator: a prognostic marker in breast cancer including patients with axillary node-negative disease. Clin Chem. 44:1177–1183. 1998.PubMed/NCBI
20.	Rath M and Pauling L: Plasmin-induced proteolysis and the role of apoprotein(a), lysine and synthetic analogs. Orthomolecular Med. 7:17–23. 1992.
21.	Andreasen PA, Kjøller L, Christensen L and Duffy MJ: The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer. 72:1–22. 1997. View Article : Google Scholar : PubMed/NCBI
22.	Niedzwiecki A, Roomi MW, Kalinovsky T and Rath M: Micronutrient synergy - a new tool in effective control of metastasis and other key mechanisms of cancer. Cancer Metastasis Rev. 29:529–543. 2010. View Article : Google Scholar : PubMed/NCBI
23.	Roebuck MM, Helliwell TR, Chaudhry IH, Kalogrianitis S, Carter S, Kemp G, Ritchie DA, Jane MJ and Frostick SP: Matrix metalloproteinase expression is related to angiogenesis and histologic grade in spindle cell soft tissue neoplasms of the extremities. Am J Clin Path. 123:405–414. 2005. View Article : Google Scholar : PubMed/NCBI
24.	Roomi MW, Monterrey JC, Kalinovsky T, Niedzwiecki A and Rath M: Inhibition of invasion and MMPs by a nutrient mixture in human cancer cell lines: a correlation study. Exp Oncol. 32:243–248. 2010.PubMed/NCBI
25.	Roomi MW, Ivanov V, Kalinovsky T, Rath M and Niedzwiecki A: In vivo and in vitro antitumor effect of ascorbic acid, lysine, proline, arginine and green tea extract on human fibrosarcoma cells HT-1080. Med Oncol. 23:105–112. 2006. View Article : Google Scholar : PubMed/NCBI
26.	Roomi MW, Ivanov V, Kalinovsky T, Niedzwiecki A and Rath M: In vitro and in vivo anti-tumor effect of a nutrient mixture containing ascorbic acid, lysine, proline and green tea extract on human synovial sarcoma cancer cells. JAMA. 9:30–34. 2006.
27.	Amin ARMR, Kucek O, Khuri FR and Shin DM: Perspectives for cancer prevention with natural compounds. J Clin Oncol. 27:2712–2725. 2009. View Article : Google Scholar : PubMed/NCBI
28.	Roomi MW, Monterrey JC, Kalinovsky T, Rath M and Niedzwiecki A: Comparative effects of EGCG, green tea and a nutrient mixture on the patterns of MMP-2 and MMP-9 expression in cancer cell lines. Oncol Rep. 24:747–757. 2010.PubMed/NCBI
29.	Sun Z, Chen YH, Wang P, Zhang J, Gurewich V, Zhang P and Liu JN: The blockage of high-affinity lysine binding sites of plasminogen by EACA significantly inhibits prourokinase-induced plasminogen activation. Biochem Biophys Acta. 1596:182–192. 2002.
30.	Kemberling JK, Hampton JA, Keck RW, Gomez MA and Selman SH: Inhibition of bladder tumor growth by the green tea derivative epigallocatechin-3-gallate. J Urol. 170:773–776. 2003. View Article : Google Scholar : PubMed/NCBI
31.	Sato D and Matsushima M: Preventive effects of urinary bladder tumors induced by N-butyl-N-(4-hydroxybutyl)-nitrosamine in rat by green tea leaves. Int J Urol. 10:160–166. 2003. View Article : Google Scholar : PubMed/NCBI
32.	Valcic S, Timmermann BN, Alberts DS, Wachter GA, Krutzsch M, Wymer J and Guillen JM: Inhibitory effect of six green tea catechins and caffeine on the growth of four selected human tumor cell lines. Anticancer Drugs. 7:461–468. 1996. View Article : Google Scholar : PubMed/NCBI
33.	Mukhtar H and Ahmed N: Tea polyphenols: prevention of cancer and optimizing health. Am J Clin Nutr. 71:1698s–1702s. 2000.PubMed/NCBI
34.	Yang GY, Liao J, Kim K, Yurtow EJ and Yang CS: Inhibition of growth and induction of apoptosis in human cancer cell lines by tea polyphenols. Carcinogenesis. 19:611–616. 1998. View Article : Google Scholar : PubMed/NCBI
35.	Taniguchi S, Fujiki H, Kobayashi H, Go H, Miyado K, Sadano H and Shimikawa R: Effect of (-) epigallocatechin gallate, the main constituent of green tea, on lung metastasis with mouse B16 melanoma cell lines. Cancer Lett. 65:51–54. 1992. View Article : Google Scholar : PubMed/NCBI
36.	Hara Y: Green tea: Health Benefits and Applications. Marcel Dekker; New York: 2001, View Article : Google Scholar
37.	Kawakami S, Kageyama Y, Fujii Y, Kihara K and Oshima H: Inhibitory effects of N-acetyl cysteine on invasion and MMP 9 production of T24 human bladder cancer cells. Anticancer Res. 21:213–219. 2001.PubMed/NCBI
38.	Morini M, Cai T, Aluigi MG, Noonan DM, Masiello L, De Floro S, D'Agostinin F, Albini A and Fassima G: The role of the thiol N-acetyl cysteine in the prevention of tumor invasion and angiogenesis. Int J Biol Markers. 14:268–271. 1999.PubMed/NCBI
39.	Yoon SO, Kim MM and Chung AS: Inhibitory effects of selenite on invasion of HT 1080 tumor cells. J Biol Chem. 276:20085–20092. 2001. View Article : Google Scholar : PubMed/NCBI
40.	Naidu KA, Karl RC and Coppola D: Antiproliferative and proapoptotic effect of ascorbyl stearate in human pancreatic cancer cells: association with decreased expression of insulin-like growth factor 1 receptor. Dig Dis Sci. 48:230–237. 2003. View Article : Google Scholar : PubMed/NCBI
41.	Anthony HM and Schorah CJ: Severe hypovitaminosis C in lung-cancer patients: The utilization of vitamin C in surgical repair and lymphocyte-related host resistance. Br J Cancer. 46:354–367. 1982. View Article : Google Scholar : PubMed/NCBI
42.	Maramag C, Menon M, Balaji KC, Reddy PG and Laxmanan S: Effect of vitamin C on prostate cancer cells in vitro: effect on cell number, viability and DNA synthesis. Prostate. 32:188–195. 1997. View Article : Google Scholar : PubMed/NCBI
43.	Koh WS, Lee SJ, Lee H, Park C, Park MH, Kim WS, Yoon SS, Park K, Hong SI, Chung MH and Park CH: Differential effects and transport kinetics of ascorbate derivatives in leukemic cell lines. Anticancer Res. 8:2487–2493. 1998.PubMed/NCBI
44.	Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, Shacter E and Levine M: Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci USA. 102:13604–13609. 2005. View Article : Google Scholar : PubMed/NCBI
45.	Nunez C, Ortiz de Apodaca Y and Ruiz A: Ascorbic acid in the plasma and blood cells of women with breast cancer. The effect of consumption of food with an elevated content of this vitamin. Nutr Hosp. 10:368–372. 1995.(In Spanish).
46.	Kurbacher CM, Wagner U, Kolster B, Andreotti PE, Krebs D and Bruckner HW: Ascorbic acid (vitamin C) improves the antineoplastic activity of doxorubicin, cisplatin and paclitaxel in human breast carcinoma cells in vitro. Cancer Lett. 103:183–189. 1996. View Article : Google Scholar : PubMed/NCBI
47.	Cooke JP and Dzau VJ: Nitric oxide synthase: role in the genesis of vascular disease. Annu Rev Med. 48:489–509. 1997. View Article : Google Scholar : PubMed/NCBI
48.	Roomi MW, Ivanov V, Netke SP, Niedzwiecki A and Rath M: Serum markers of the liver, heart, and kidney and lipid profile and histopathology in ODS rats treated with nutrient synergy. J Am Coll Nutr. 22:477abst 86,. 2003.