Inhibition of the SK-N-MC human neuroblastoma cell line in vivo and in vitro by a novel nutrient mixture

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

doi:10.3892/or.2013.2307

Inhibition of the SK-N-MC human neuroblastoma cell line in vivo and in vitro by a novel nutrient mixture

Authors:
- M. Waheed Roomi
- Tatiana Kalinovsky
- Nusrath W. Roomi
- Aleksandra Niedzwiecki
- Matthias Rath
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Affiliations: Dr Rath Research Institute, Oncology Division, Santa Clara, CA, USA
Published online on: February 27, 2013 https://doi.org/10.3892/or.2013.2307
Pages: 1714-1720
Copyright: © Waheed 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

Neuroblastoma, a peripheral nervous system cancer that can be highly invasive and metastatic, accounts for 8-10% of all solid childhood tumors in children under the age of 15 years. Despite multiple clinical efforts, prognosis remains poor for this enigmatic disease. A nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has shown significant antitumor effects. Using the nude mouse xenograft model, we investigated the efficacy of NM. We also tested the effect of NM in vitro, evaluating cell viability, secretion of the matrix metalloproteinases (MMP)-2 and MMP-9, tissue inhibitor of metalloproteinase (TIMP)-2 secretion, Matrigel invasion and cellular apoptosis and morphology. Athymic nude mice 5-6 weeks of age were inoculated with 3x106 SK-N-MC neuroblastoma cells subcutaneously and randomly divided into two groups. Group A was fed a regular diet and group B a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighed and processed for histology. We also tested the effect of NM in vitro. NM inhibited the growth of xenograft tumors by 22% (P=0.04); and, in vitro, NM induced dose-dependent inhibition of cell proliferation with a decrease of 27% (P=0.001) and 36% (P=0.002) at 500 and 1000 µg/ml NM compared to the control, respectively. Zymography revealed MMP-2 secretion in normal cells and PMA (100 ng/ml)‑induced MMP-9 secretion. NM inhibited the secretion of both MMPs with total blockage at a concentration of 100 µg/ml. Reverse zymography demonstrated a dose-dependent increase in TIMP-2 expression by NM. Notable, SK-N-MC human neuroblastoma cells were not invasive through Matrigel. NM induced dose-dependent apoptosis of SK-N-MC cells. The results suggest that NM may have therapeutic potential in treating neuroblastoma.

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1	American Cancer Society. Neuroblastoma: What are the key statistics about neuroblastoma? http://www.cancer.org/Cancer/Neuroblastoma/DetailedGuide/neuroblastoma-key-statistics. Accessed Dec 6, 2012
2	Reynolds CP and Seeger RC: Neuroblastoma. Cancer Treatment. Haskell CM: W.B. Saunders; Philadelphia, PA: pp. 860–871. 1994
3	Matthay KK, Villablanca JG, Seeger RC, Stram DO, Harris RE, Ramsay NK, Swift P, Shimada H, Black CT, Brodeur GM, Gerbing RB and Reynolds CP: Treatment of high-risk neuroblastoma with intensive chemotherapy, radiotherapy, autologous bone marrow transplantation and 13-cis-retinoic acid. Children’s Cancer Group. N Engl J Med. 341:1165–1173. 1999.
4	Roy Choudhury S, Karmakar S, Banik NL and Ray SK: Targeting angiogenesis for controlling neuroblastoma. J Oncol. 2012:7820202012. View Article : Google Scholar
5	Ara T, Kusafuka T, Inoue M, Kuroda S, Fukuzawa M and Okada A: Determination of imbalance between MMP-2 and TIMP-2 in human neuroblastoma by reverse-transcription polymerase chain reaction and its correlation with tumor progression. J Pediatr Surg. 35:432–437. 2000. View Article : Google Scholar : PubMed/NCBI
6	Sugiura Y, Shimada H, Seeger RC, Laug WE and DeClerck YA: Matrix metalloproteinases-2 and-9 are expressed in human neuroblastoma: contribution of stromal cells to their production and correlation with metastasis. Cancer Res. 58:2209–2216. 1998.
7	Roomi MW, Roomi N, Ivanov V, Kalinovsky T, Niedzwiecki A and Rath M: Inhibitory effect of a mixture containing ascorbic acid, lysine, proline and green tea extract on critical parameters in angiogenesis. Oncol Rep. 14:807–815. 2005.PubMed/NCBI
8	Roomi MW, Roomi N, Ivanov V, Kalinovsky T, Niedzwiecki A and Rath M: Inhibition of pulmonary metastasis of melanoma B16FO cells in C57BL/6 mice by a nutrient mixture consisting of ascorbic acid, lysine, proline, arginine, and green tea extract. Exp Lung Res. 32:517–530. 2006. View Article : Google Scholar : PubMed/NCBI
9	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. 29:529–542. 2010. View Article : Google Scholar : PubMed/NCBI
10	Ribatti D, Marimpietri D, Pastorino F, Brignole C, Nico B, Vacca A and Ponzoni M: Angiogenesis in neuroblastoma. Ann NY Acad Sci. 1028:133–142. 2004. View Article : Google Scholar
11	Ribatti D, Vacca A, Nico B, De Falco G, Montaldo GP and Pnzoni M: Angiogenesis and anti-angiogenesis in neuroblastoma. Eur J Cancer. 38:750–757. 2002. View Article : Google Scholar : PubMed/NCBI
12	Rath M and Pauling L: Plasmin-induced proteolysis and the role of apoprotein(a), lysine and synthetic analogs. J Orthomolecular Med. 7:17–23. 1992.
13	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.PubMed/NCBI
14	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
15	Mukhtar H and Ahmad N: Tea polyphenols: prevention of cancer and optimizing health. Am J Clin Nutr. 71(Suppl 6): S1698–S1704. 2000.PubMed/NCBI
16	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
17	Taniguchi S, Fujiki H, Kobayashi H, Go H, Miyado K, Sadano H and Shimokawa 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.
18	Hara Y: Green Tea: Health Benefits and Applications. Marcel Dekker, Inc; New York, Basel: 2001, View Article : Google Scholar
19	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
20	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
21	Yoon SO, Kim MM and Chung AS: Inhibitory effects of selenite on invasion of HT1080 tumor cells. J Biol Chem. 276:20085–20092. 2001. View Article : Google Scholar : PubMed/NCBI
22	Carosio R, Zuccari G, Orienti I, Mangraviti S and Montaldo PG: Sodium ascorbate induces apoptosis in neuroblastoma cell lines by interfering with iron uptake. Mol Cancer. 6:552007. View Article : Google Scholar : PubMed/NCBI
23	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
24	Naidu KA, Karl RC, Naidu KA 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
25	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. 18:2487–2493. 1998.PubMed/NCBI
26	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
27	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
28	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
29	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).
30	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
31	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. 22:4772003.