Background: Matrix metalloproteinases (MMPs), vascular endothelial growth factor (VEGF), Ki 67 (proliferative protein) and
constituents of ECM play a critical role in angiogenesis, and are crucial in ...neoplastic invasion and metastasis. Based on
the antitumor properties of certain nutrients, we investigated the effect of a diet containing lysine, proline, arginine,
ascorbic acid and green tea extract on the growth of tumors induced by implanting human prostate cancer PC-3 cells in athymic
nude mice and on the expression of MMPs, VEGF, Ki 67 and fibronectin in these tumors, as well as the production of mucin (by
PAS staining). Materials and Methods: Male nude mice (n=12) were inoculated with 3Ã10 6 prostate cancer PC-3 cells and randomly divided into two groups; Group A was fed a regular diet and Group B was fed a regular
diet supplemented with 0.5% of the nutrient mixture (NM). Four weeks later, tumors were excised, weighed and processed for
histology. Results: The results showed inhibition of tumor growth in Group B. Histological studies revealed inhibition of
MMP-9 and VEGF secretion and mitosis in Group B tissues. Conclusion: Nutrient supplementation strongly suppressed the growth
of tumors without any adverse effects in nude mice, suggesting strong potential as an anticancer agent.
Current treatment of osteosarcoma is associated with poor prognosis, especially due to the increased risk of developing other cancers with chemotherapy. Therefore, new, safe and effective treatment ...strategies are needed. We investigated the effect of a unique mixture of nutrients containing lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate (EGCG) on human osteosarcoma cell lines U-2OS, MNNG-HOS, and Ewing's sarcoma SK-ES-1 by measuring: cell proliferation, expression of matrix metalloproteinase-2 (MMP-2), MMP-9, and invasive and angiogenesis potential. Cell proliferation was evaluated by MTT assay, matrix metalloproteinases (MMP) expression by gelatinase zymography, VEGF expression by ELISA, and invasion through Matrigel. Cells were also treated with phorbol 12-myristate 13-acetate (PMA) to study enhanced MMP and VEGF expression. The invasion of osteosarcoma U-2OS and MNNG-HOS cells through Matrigel was significantly reduced in a dose-dependent fashion, with 100% inhibition of invasion of U-2OS cells at 100 microg/ml, and MNNG cells at 50 microg/ml concentration of the synergistically acting nutrient mixture. Ewing's sarcoma SK-ES-1 cells were not invasive. Nutrient synergy (NS) exhibited a dose response antiproliferative effect on osteosarcoma U-2OS cells, reaching 67% at 1000 microg/ml of NS; no significant suppression of cell proliferation was seen with MNNG or Ewing's sarcoma cells. Zymography showed dose-dependent inhibition of MMP secretion by all three cell lines in the presence of NS. VEGF secretion by U-2OS cells was completely blocked at 500 microg/ml of NS. Our results suggest NS is an excellent candidate for therapeutic use in the treatment of osteosarcoma, by inhibiting cancer cell invasion, and secretion of MMPs and VEGF, all critical parameters for cancer control and prevention.
Abstract
Breast cancer patients often have detectable or occult metastases at diagnosis and most patients will develop metastatic lesions during the course of the disease. We investigated the effect ...of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract on murine breast cancer 4T1, a unique metastatic breast cancer model that has the capacity to metastasize efficiently to sites affected in human breast cancer. After one week of isolation, 5-6 week old female Balb/C mice were inoculated with 5x105 4T1 cells into the mammary pad and randomly divided into two groups; group A was fed a regular diet and group B a regular diet supplemented with 0.5% NM. After four weeks, the mice were sacrificed and their tumors, lungs, livers, kidneys, hearts and spleens were excised and processed for histology. Dimensions (length and width) of tumors were measured using a digital caliper, and the tumor burden was calculated using the following formula: 0.5 x length x width. We also tested the effect of NM in vitro on 4T1 cells, measuring cell proliferation by MTT assay, MMP secretion by zymography, invasion through Matrigel , migration by scratch test and morphology by H&E staining. NM inhibited tumor weight and burden of 4T1 tumors by 50% (p =0.02) and 53.4% (p<0.0001), respectively. Lung metastasis was profoundly inhibited by NM supplementation: mean number of colonies was reduced by 87% (p<0.0001) and mean weight of lungs by 60% (p=0.0001) compared to control mice. Metastasis to liver, spleen, kidney and heart was significantly reduced with NM supplementation. In vitro, NM exhibited 50% toxicity over the control at 250 and 500 µg/ml concentrations. Zymography demonstrated MMP-2 and MMP-9 secretion which was inhibited by NM in a dose dependent fashion, with virtual total inhibition of both at 1000 µg/ml. Migration by scratch test and Invasion through Matrigel were inhibited in a dose dependent manner with total block of invasion at 250 µg/ml and of migration at 1000 µg/ml. These results suggest that NM has therapeutic potential in treatment of breast cancer.
Citation Format: M. Waheed Roomi, John Cha, Nusrath Roomi, Aleksandra Niedzwiecki, Matthias Rath. In vivo and in vitro effect of a nutrient mixture on murine 4T1 breast cancer. abstract. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4963. doi:10.1158/1538-7445.AM2014-4963
Standard multimodality therapy of gliomas is associated with poor patient survival and significant toxicity. Abnormal expression of matrix metalloproteinases is associated with tumor growth and ...invasion. Based on reported antitumor properties, we investigated the effect of a combination of natural compounds (NM), primarily composed of lysine, proline, ascorbic acid, and green tea extract in vitro on glioma cell line A-172, by measuring MMP secretion, invasion through Matrigel, and cell proliferation. Glioma cells A-172 (ATCC) were grown in modified Dulbecco's Eagle medium with 10% fetal bovine serum and antibiotics and treated with NM at 0, 10, 50, 100, 500, and 1000 microg/mL concentration in triplicate at each dose. Cell proliferation was assayed by MTT, MMP secretion by zymography, invasion through Matrigel, and morphology by H&E staining. Zymography showed one band corresponding to MMP-2, which was inhibited by NM in a dose-dependent fashion, with virtual total inhibition at 500-microg/mL concentration. Invasion through Matrigel was completely inhibited at 1000 microg/mL NM. NM was not toxic to glioma cell line A-172 at lower concentrations and exhibited toxicity of 50% over the control at 1000 microg/mL. NM significantly inhibited MMP secretion and invasion-important parameters for cancer prevention, suggesting a possible therapeutic role.
The high incidence of lung cancer and ineffective toxic action of current mono and doublet chemotherapy approaches result in poor patient survival. Further, matrix metalloproteinases (MMPs) are ...implicated in neoplastic invasion and metastasis. Based on this, the authors investigated the effect of a dietary micronutrient mixture (NM) containing lysine, proline, arginine, ascorbic acid, and green tea extract on the tumor growth of human lung carcinoma cell A-549 xenografts in athymic nude mice. Additionally, the authors tested the in vitro antitumor effect of NM on lung carcinoma A-549 cells by measuring cell proliferation by MTT assay, MMP-2 and -9 secretion by gelatinase zymography, and cell invasion through Matrigel. Nutrient supplementation strongly suppressed the growth of tumors without adverse effects in nude mice; tumor weight was reduced by 44% (P = .0001) and tumor burden was reduced by 47% (P < .0001) with supplementation. Zymography demonstrated in vitro secretion of MMP-2 by uninduced human lung carcinoma cells and both MMP-2 and -9 by phorbol 12-mysristate 13-acetate (PMA) (200 ng/mL)-treated cells. NM inhibited the secretion of both MMPs in a dose-dependent fashion, with virtual total inhibition at 500 µg/mL concentration. The invasion of human lung carcinoma cells through Matrigel was significantly reduced at 100 µg/mL (64%) and totally inhibited at 500 µg/mL concentration of NM (P = .01). Suppression of lung tumor growth in nude mice and inhibition of MMP secretion and Matrigel invasion suggest NM may act as an anticancer agent and as such warrants further investigation.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
Testicular cancer (TC) is rare, but still represents one of the most common diseases in young men between the ages of 20-45. However, men of any age can develop this disease. The incidence ...in Caucasians is greater than in African Americans. Risk factors include undescended testis, Klinefelter syndrome, and HIV positive patients. If left untreated, it is almost certainly fatal. Metastasis is the major cause of cancer death. The most common place for TC to spread in the body is to the lung. In this investigation, we studied the effect of a novel nutrient mixture (NM) containing ascorbic acid, amino acids and green tea extract that has been shown to exhibit anti-cancer activity, on inhibition of B16FO melanoma cells inoculated intratesticularly. Male athymic mice (n=12), 10-12 weeks of age, were inoculated with half a million B16FO melanoma cells in 100 µL of PBS into the right testis; the left testis was left untreated. After inoculation, the mice were randomly divided into two groups. Group A (n=6) was fed a regular mouse chow diet, while the mice in Group B (n=6) were fed the same diet but supplemented with 0.5% NM. Four weeks later the mice were sacrificed and the abdominal cavity was opened. Mice in the control group (Group A) exhibited extensive metastasis in the peritoneal cavity, which was totally masked by B16FO melanoma cells. The testis was severely enlarged and replaced by invading malignant melanoma cells. The remaining testicular tissue was represented by necrotic seminiferous tubules. The capsular region of the testis was severely infiltrated with a population of mixed cells. In contrast, in the NM fed group (Group B), there was no evidence of peritoneal metastasis, but the testes were enlarged. Seminiferous tubules in the area of invasion showed evidence of degeneration. In all groups, there was no metastasis to liver, kidney and spleen. However, severe lung metastasis was observed in the control group (2 out of 6) and mild in the test group (2 out of 6). In conclusion, these results suggest that NM has potential to suppress tumor metastasis.
Citation Format: {Authors}. {Abstract title} abstract. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2348. doi:1538-7445.AM2012-2348
Background: Melanoma, a very serious form of skin cancer, causes the most skin cancer-related deaths, due to metastasis. Structural
changes in the extracellular matrix (ECM) are necessary for cell ...migration during tissue remodeling. MMPs, VEGF, Ki-67 (proliferative
protein) and constituents of ECM play a critical role in angiogenesis, and are crucial in neoplastic invasion and metastasis.
Materials and Methods: The effect of a diet (NM) containing lysine, proline, arginine, ascorbic acid and green tea extract
on the growth of tumors induced by implanting human melanoma A2058 cells in athymic nude mice was examined and, also, on the
expression of MMPs, VEGF and Ki-67 in these tumors. The effect of NM in vitro on the melanoma A2058 cell line was tested by
measuring: cell proliferation by the MTT assay, expression of MMPs by gelatinase zymography and invasion through Matrigel.
Results: Nutrient supplementation strongly suppressed the growth of tumors (by 57%)without adverse effects in nude mice. Histological
studies supported these findings by showing inhibition of MMP-9 and VEGF secretion and mitotic index. In vitro, NM inhibited
melanoma cell growth by 64% at 500 μg/ml and Matrigel invasion by 95% at 100 μg/ml NM. Conclusion: These results suggest that
NM may have a therapeutic potential in melanoma.
Matrix metalloproteinases (MMPs) have been recognized as key players in the degradation of the extracellular matrix (ECM) by migration and proliferation of endothelial cells and their subsequent ...invasion of the underlying stroma. The prevention of ECM degradation through the inhibition of MMP activity has been shown to be a promising therapeutic approach to block the invasion that occurs during angiogenesis. In previous studies, we demonstrated the anti-tumor effect of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, green tea extract, arginine, N-acetyl cysteine, selenium, copper and manganese on various tumor cell lines in vivo and in vitro. The aim of the present study was to determine whether this mixture has anti-angiogenic effects on human umbilical vein endothelial cells (HUVECs). At near confluence, the HUVEC cell cultures were tested with NM at 0, 10, 50, 100, 500, and 1000 microg/ml in triplicate at each dose for proliferation, migration, MMP expression, and invasion. Cell proliferation was evaluated by MTT assay, invasion potential by Matrigel invasion, MMP expression by gelatinase zymography, and cell migration by a 2 mm-wide scratch in plates. For tube formation, HUVECs were cultured in previously polymerized Matrigel. NM inhibited HUVEC migration, MMP expression and invasion through Matrigel in a dose-dependent manner. Zymography showed a dose-dependent inhibition of MMP-2 expression with virtual total inhibition at a 500 microg/ml concentration. Invasion through Matrigel was totally inhibited at 500 microg/ml NM. NM reduced cell migration by scratch test in a dose-dependent fashion with total inhibition at a 500 microg/ml concentration. NM also inhibited the tube formation of HUVECs, but did not significantly inhibit cell proliferation. These results together with our earlier findings suggest that NM is a relatively non-toxic formulation with anti-angiogenic effects, such as inhibiting vascular tube formation and endothelial cell invasion and migration.
Abstract
Our main objective was to determine the differential response of female gulonolactone oxidase (Gulo) mice challenged with 4T1 to vitamin C deficient, subclinical vitamin C sufficient, or ...vitamin C combined with other additional natural products (EQC) diets against 4T1 secondary growth phase metastasis to lungs, kidneys, and liver. Live murine 4T1 cells (5x105) were administered subcutaneously to the right flank of female Gulo (-/-) mice (n=27) 35-40 weeks of age and 18 age matched female wild-type mice. Tumors were established for an additional 2 weeks on subclinical vitamin C (100mg/L in water) in the Gulo (-/-) group, or with regular food and water in the wild-type vitamin C-generating mice. Tumor measurements were taken and the Gulo (-/-) mice were distributed into 3 groups (n=9 in each) to ensure average tumor burden was equivalent before therapy. Mice were then maintained for an additional 2 weeks on either subclinical vitamin C in water, subclinical C water + vitamin C (equivalent to that provided by 0.5% EQC supplemented diet), and subclinical C water + 0.5% EQC supplemented diet. Wild-type mice received an additional 2 weeks of either regular murine diet or 0.5% EQC supplemented food and regular water. At the endpoint, serum was collected, lungs, livers, and kidneys were evaluated for metastatic burden, and tissues collected for histology. Tumors were also harvested, weighed, and sectioned for histology. EQC exhibited the strongest anti-metastatic effect upon the second growth phase in scorbutic Gulo (-/-) mice as well as wild type vitamin C generating mice. Gulo (-/-) mice maintained throughout the study on subclinical vitamin C presented with the worst outcomes, including novel renal involvement. Vitamin C in diet equivalent to that in 0.5% EQC afforded only protection against novel renal pathology, but the metastatic burden was similar to that in subclinical C group. 0.5% EQC abrogated the cases of moderate to severe metastasis in scorbutic Gulo (-/-) mice by 16% compared to subclinical vitamin C group. In wild-type mice, 0.5% EQC abrogated the cases of moderate to severe metastasis by 37%. This study demonstrates that 0.5% EQC is superior to vitamin C alone in reducing metastasis from a primary 4T1 tumor and that prior scurvy is deleterious upon host resistance to primary tumors and reduces the efficacy of subsequent therapy against metastasis. Prior endogenous vitamin C generation in wild-type mice before tumor inoculation and continuous vitamin C generation during therapy with 0.5% EQC suggests that vitamin C both enhances host resistance and synergistically potentiates the effect of additional therapy. Tumor mass itself was not related to metastatic burden, suggesting that differing biochemical composition of the stromal and tumor structures are responsible for a metastatic or non-metastatic tumor of equal size.
Citation Format: John Cha, M. Waheed Roomi, Matthias Rath, Aleksandra Niedzwiecki. Ascorbic acid synergistically potentiates antimetastatic effect of natural products on 4T1 tumors. abstract. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2822. doi:10.1158/1538-7445.AM2013-2822
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