A Specific Mixture Of Nutrients Suppresses Ovarian Cancer A-2780 Tumor Incidence, Growth, And Metastasis To Lungs

M. Waheed Roomi, Tatiana Kalinovsky, Matthias Rath, Aleksandra Niedzwiecki
Dr. Rath Research Institute, Santa Clara, California, USA
Nutrients  9:303, 2017

Abstract
Ovarian cancer is the deadliest gynecological malignancy in women, and fifth leading cause of death. Despite advances made in chemotherapy and surgery, the average time of clinical remission is approximately 2 years and the 5-year survival rate is 45%. Thus, there is an urgent need for the development of a novel therapeutic approach to ovarian cancer treatment. We investigated the effect of a specific nutrient mixture (EPQ) containing ascorbic acid, lysine, proline, green tea extract, and quercetin on human ovarian cancer cell A-2780 in vivo and in vitro. Athymic female nude mice (n = 12) were all inoculated intraperitoneally (IP) with 2 x106 cells in 0.1 mL of phosphate buffered saline (PBS) and randomly divided into two groups. Upon injection, the Control group (n = 6) was fed a regular diet and the EPQ group (n = 6) a regular diet supplemented with 0.5% EPQ. Four weeks later, the mice were sacrificed and tumors that developed in the ovary were excised, weighed, and processed for histology. Lungs were inspected for metastasis. In vitro, A-2780 cells were cultured in Dulbecco modified Eagle medium supplemented with 10% FBS and antibiotics. At near confluence, cells were treated with EPQ in triplicate at concentrations between 0 and 1000 μg/mL. Cell proliferation was measured via MTT assay, MMP-9 secretion via gelatinase zymography, invasion through Matrigel and morphology via hematoxylin and eosin (H & E) staining. All Control mice developed large ovarian tumors, whereas 5 out of 6 mice in the EPQ group developed no tumors, and one, a small tumor. Control mice also showed lung metastasis in 6 out of 6 mice, while no lung metastasis was evident in EPQ mice. Zymography demonstrated only MMP-9 expression, which EPQ inhibited in a dose-dependent fashion, with virtual total block at 250 μg/mL concentration .EPQ significantly inhibited invasion through Matrigel with total block at 250 μg/mL concentration. MTT showed dose-dependent inhibition of cell proliferation with EPQ, and H & E staining showed no morphological changes below 500 μg/mL EPQ. These results suggest that EPQ has therapeutic potential in the treatment of ovarian cancer by significantly suppressing ovarian tumor incidence and growth and lung metastasis, and by inhibiting MMP-9 secretion and invasion of A-2780 ovarian cancer cells.

Keywords: ovarian cancer A-2780 cell line; nutrients; tumor growth; MMPs; Matrigel invasion;
metastasis

Access: http://www.mdpi.com/2072-6643/9/3/303

A Nutrient Mixture Modulates Ovarian ES-2 Cancer Progression By Inhibiting Xenograft Tumor Growth And Cellular MMP Secretion, Migration And Invasion

M.W. Roomi, T. Kalinovsky, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
International Journal of Clinical and Experimental Medicine 9(2):814-822, 2016

Abstract:

Epithelial ovarian carcinoma, which occurs mainly in post-menopausal women, is the leading cause of death from gynecological malignancy and the fifth most common cancer in the U.S. Since ovarian cancer often remains clinically silent, the majority of patients with ovarian carcinoma have advanced intraperitoneal metastatic disease at diagnosis, resulting in a poor prognosis. Long-term survival of patients with ovarian cancer remains poor, due to metastasis and recurrence.

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Effect of a nutrient mixture on the localization of extracellular matrix proteins in HeLa human cervical cancer xenografts in female nude mice

M.W. Roomi, T. Kalinovsky,  A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
International Journal of Oncology 2015 DOI: 10.3892/ijo.2015.3008

Abstract:

Colorectal, pancreatic and hepatic carcinomas 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 uPA and MMPs with cancer progression, metastasis and shortened patient survival.

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Effects of a nutrient mixture on immunohistochemical localization of cancer markers in human cervical cancer HeLa cell tumor xenografts in female nude mice

M.W. Roomi, T. Kalinovsky, J. Cha, N.M. Roomi,  A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
Experimental & Therapeutic Medicine 2014, DOI: 10.3892/etm.2014.2127

Abstract:
Although fully treatable in the early stages, once cervical cancer has metastasized, patient outcome is poor. The main objective of this study was to examine the effect of dietary supplementation with a nutrient mixture (NM) containing lysine, ascorbic acid, proline, green tea extract and other micronutrients on HeLa cell xenografts in nude female mice. Tumor growth was measured and immunohistochemical staining was evaluated for the following cancer markers: Ki67 (proliferation); matrix metalloproteinase (MMP) 2 and  9 (invasion/metastasis); vascular endothelial growth factor (VEGF) (angiogenesis); terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and B-cell lymphoma 2 (Bcl-2) (apoptosis); cyclooxygenase 2 (COX 2) and inducible nitric oxide synthase (iNOS) (inflammation); and glutathione S-transferase π (GSTπ) (a general cancer marker).

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

M.W. Roomi, T. Kalinovsky, M. Rath and A. Niedzwiecki
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
Oncology Reports 2012, 28:768-776

Abstract:
Cancers of the breast, cervix, uterus and ovary are the most prevalent cancers in women worldwide. 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 urokinase plasminogen activators (u-PA) and matrix metalloproteinases (MMPs) with cancer progression, metastasis and shortened patient survival. MMP activities are regulated by specific tissue inhibitors of metalloproteinases (TIMPS).

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Anticancer Effects of a Specific Mixture of Nutrients in Multidrug Resistant Human Uterine Sarcoma Cell Line MES-SA/Dx5 and in Drug Sensitive MES-SA Cell Line

 

M.W. Roomi, T. Kalinovsky, N.W. Roomi, M. Rath and A. Niedzwiecki
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050
Oncology Reports 2011, 27: 17-27

Abstract:
A specific nutrient mixture containing lysine, proline, ascorbic acid and green tea extract (NM) has demonstrated a broad spectrum of antitumor activity against a number of cancer cell lines. In this study, our main objective was to investigate the comparative effects of NM on anticancer parameters such as cytotoxicity, MMP secretion, and Matrigel invasion in the human uterine sarcoma drug-resistant MES-SA/Dx5 and drug sensitive drug-sensitive MES-SA cell lines. In addition we studied the effect of NM on P-glycoprotein (Pgp) on these cell lines.

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In Vitro Modulation of MMP-2 and MMP-9 in Human Cervical and Ovarian Cancer Cell Lines by Cytokines, Inducers, and Inhibitors

M.W. Roomi, J.C. Monterrey, T. Kalinovsky, M. Rath, A. Niedzwiecki
Oncology Reports 2010, 23: 605-614

Abstract:
Matrix  metalloproteinases (MMPs) secreted by cervical and ovarian cancer, especially  MMP-2 and MMP-9, play crucial roles in tumor invasion and metastasis. We examined the effect of cytokines, mitogens, inducers and inhibitors on MMP-2 and MMP-9 expression in cervical and ovarian cancer cell lines. Human cervical (Hela and DoTc2-4510) and ovarian (SK-OV-3) cell lines were cultured in appropriate media. At near confluence, the cells were washed with PBS and incubated in serum-free medium with various concentrations of several cytokines, mitogens and inhibitors. After 24h the media were removed and analyzed for MMP-2 and MMP-9 by gelatinase zymography and quantitated by densitometry.

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Suppression of Human Cervical Cancer Cell Lines Hela and DoTc2 4510 MMP Expression and Matrigel Invasion by a mixture of lysine, proline, ascorbic acid, and green tea extract

M.W. Roomi, V. Ivanonv, T. Kalinovsky, A. Niedzwiecki, M. Rath
International Journal of Gynecological Cancer 2006, 16(3): 1241-1247

Introduction:
Cervical cancer, the second most common cancer in women, once metastasized, leads to poor prognosis. We investigated the antitumor effect of a mixture (NM) containing lysine, proline, arginine, ascorbic acid, and green tea extract on human cervical cancer cells Hela (CCL2) and DoTc2 4510.

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Inhibition of Matrix Metalloprotenase-2 Secretion and Invasion by Human Ovarian Cancer Cell Line SK-OV-3 with Lysine, Proline, Ascorbic Acid, and Green Tea Extract

M.W. Roomi, V. Ivanov, T. Kalinovsky, A. Niedzwiecki, M. Rath
Journal of Obstetrics and Gynaecology Research 2006, 32(2):148-154.

Introduction:
Based on the poor prognosis associated with ovarian cancer and reported anti-cancer properties of specific nutrients, we investigated the effect of a nutrient mixture (NM) containing lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate on human ovarian cancer cells SK-OV-3 by measuring: cell proliferation, modulation of MMP-2 and –9 secretion, and cancer cell invasive potential.

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