Scientific Presentations

Scientific Presentations: Cancer

Marked Inhibition of Tumor Growth, MMP Secretion and Invasion by a Nutrient Mixture on Head and Neck Squamous Carcinoma Cell Line FaDu: In Vitro and In Vivo Studies

M.W. Roomi, V. Ivanov, A. Niedzwiecki, M. Rath
Dr. Rath Research Institute, Oncology, 1260 Memorex Drive, Santa Clara, CA 95050

Presented at:
18th EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics, November 7-10, 2006, Prague, Czechoslovakia

Published in: 
The European Journal of Cancer 2006, vol 4(12), abstract #520.

Abstract

Background:
Head and neck squamous cell carcinomas (HNSCC), the sixth most common malignancies in the United States, are known for their aggressive growth and propensity to invade and metastasize. We investigated the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract on human HNSCC cell line FaDu in vitro, evaluating viability, MMP secretion, invasion and morphology. In vivo studies were carried out in athymic nude mice bearing HNSCC FaDu xenografts.

Methods:
After one week of isolation, 5-6 weeks old athymic male nude mice were inoculated with 3x106 FADU 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 on FaDu cells, measuring cell proliferation by MTT assay, invasion through Matrigel, morphology by H&E staining, and secretion of MMPs by gelatinase zymography. Cells were also treated with PMA for MMP-9 induction.

Results:
NM strongly inhibited the growth of tumors by 50%. In vitro, NM exhibited dose response toxicity with maximum toxicity of 50% over the control at 1000 mg/ml. Zymography showed only a faint band representing MMP-2 and PMA-induced MMP-9. NM inhibited secretion of both MMP-2 and MMP-9 in a dose dependent fashion, with virtual total inhibition at 1000 mg/ml. Invasion through Matrigel was inhibited at 50, 100 and 500 mg/ml by 75%, 85% and 100% respectively. H&E staining did not indicate changes up to 100 µg/ml.

Conclusion:
In conclusion, NM has a great potential for therapeutic use in the treatment of HNSCC by suppressing tumor growth and significantly inhibiting MMP secretion and invasion of HNSCC cells in vitro.

Comment

Head and neck squamous cell carcinomas (HNSCC), the sixth most common malignancies in the United States, are known for their aggressive growth and propensity to invade and metastasize. We investigated the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract, shown to have potent antitumor effects on many cancer cell lines, on the growth of human HNSCC xenografts in athymic nude mice, as well as investigating this cell line in vitro, evaluating viability, MMP secretion, invasion and morphology. NM inhibited the growth of tumors by 50% and exhibited dose-response toxicity to cells in vitro, with 50% toxicity at 1000 µg/ml. In addition, NM inhibited the invasive parameters of MMP secretion and Matrigel invasion by cells in a dose-dependent manner with total block of MMPs at 1000 µg/ml and invasion at 500 µg/ml. These results are significant as they suggest that NM, a relatively safe therapeutic agent, has great potential in treatment of HNSCC in suppressing tumor growth and metastasis.


Suppression of Growth In Vivo and In Vitro of Murine B16FO Melanoma Cells by a Novel Nutrient Mixture

M.W. Roomi, V. Ivanov, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, Oncology Division, 1260 Memorex Drive, Santa Clara, CA 95050

Presented at: 
46th Annual Meeting of The American Society for Cell Biology, San Diego December 9-13, 2006.

Published in: 
46th Annual Meeting of The American Society for Cell Biology, San Diego, December 9-13, 2006 Proceedings, Abstract # 1280.

Abstract

Introduction:
A novel nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has exhibited anti-tumor activity in vivo and vitro. In this study we examined the effect of NM on melanogenesis in vivo and in vitro using B16-F0 melanoma cell line. In advanced stages, highly metastatic melanoma is resistant to existing therapies.

Objective:
We investigated the effect of NM on murine B16FO melanoma cells in vitro evaluating viability, MMP secretion, invasion, morphology and apoptosis. In vivo studies were carried out in athymic nude mice bearing B16-F0 xenografts.

Methods:
Athymic nude male mice, 5-6 weeks old, were inoculated with 1x106 B16-FO melanoma cells (ATCC) 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, measuring cell proliferation by MTT assay, invasion through Matrigel, secretion of MMPs by gelatinase zymography, cell morphology by H&E staining and apoptosis using live green caspase detection kit (Molecular Probes).

Results:
NM inhibited the growth of B16-FO melanoma cells in vivo by 50%. Lesions, both in control and test groups were composed of cords and nests of large, irregularly round, pigmented cells consistent with a malignant melanoma. In vitro, NM was not toxic to the melanoma cells at 100 mg/ml concentration, but exhibited 50% toxicity over the control at 500 and 1000 mg/ml. H&E did not indicate any morphological changes up to 100 mg/ml. B16-F0 melanoma cells demonstrated no MMP secretion nor invasion through Matrigel. NM induced slight apoptosis at 100 mg/ml, moderate at 500 and extensive at 1000 mg/ml concentration.

Conclusion:
Taken together these results suggest that NM has many attractive features as a new anti-tumor agent.

Comment

Melanoma is a very serious and highly metastatic form of skin cancer, which causes the most skin cancer-related deaths. In its advanced stages melanoma is resistant to existing therapies. We investigated the effect of a unique nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract on murine B16FO melanoma cells in vitro and also in vivo by injecting these melanoma cells under the skin of nude mice. After 4 weeks of supplementation with NM growth of melanoma tumors in mice was inhibited by 50%. Melanoma cells exposed to 500 and 1000 mg/ml NM concentration in vitro, exhibited 50% toxicity over the control. At these concentrations a moderate and extensive apoptosis (natural cell death) was observed respectively. These results are significant as they suggest NM as a therapeutic agent for melanoma.


Apoptosis of Human Cervical and Ovarian Cancer Cell Lines Induced by a Novel Nutrient Mixture

M.W. Roomi, N.W. Roomi, V. Ivanov, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050

Presented at: 
GTCbio Apoptosis in Drug Discovery; San Diego, CA; March 22-23, 2007

Published in: 
Proceedings of the GTCbio Conference on Apoptosis in Drug Discovery

Abstract

Introduction: 
Cervical cancer is the fourth most common malignant neoplasm and ovarian cancer the seventh most common. Once these cancers are metastasized, 5-year survival is limited to 10-20%. A unique antitumor agent, a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has demonstrated anticarcinogenic activity against a number of human cancer cell lines.

Objective: 
We investigated whether the antitumor effect of NM on cervix and ovarian cancer cell lines was due to apoptosis.

Method and Materials: 
Cervix (Hela) and ovarian (SK-OV-3) cancer cell lines from ATCC were cultured in MEM and McCoy medium and treated with NM in different doses, 0, 100, 500 and 1000 µg/ml. Growth inhibition was detected by MTT assay, morphology by H&E staining, and apoptosis by Live Green Caspase Detection Kit.

Results: 
NM was slightly toxic to Hela cells at 100 µg/ml and significant at 500 and 1000 µg/ml. In contrast, NM showed slight insignificant effect on ovarian cancer cell growth. H&E staining revealed a similar apoptotic pattern in Hela and SK-OV-3 cells treated with NM at 100, 500 and 1000 µg/ml: cell shrinkage, condensation of nuclei, and acidophilic cytoplasm. Using the caspase kit, both early and late apoptosis were detected in Hela and ovarian cells: moderate at 100 µg/ml NM and significant at 1000 µg/ml NM.

Conclusions: 
The results suggest that NM induces apoptosis in both cervical and ovarian cancer cell lines and thus as a promising new therapeutic agent.

Comment

A unique antitumor agent, a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has demonstrated anticarcinogenic activity against a number of human cancer cell lines, including ovarian and cervical cancer. We investigated whether the antitumor effect of NM on cervical and ovarian cancer cell lines was due to apoptosis. NM was toxic to cervical (Hela) cells but did not affect ovarian cancer (SK-OV-3) cells. However, morphology and caspase apoptosis assays indicated that NM induced apoptosis in both cells lines – moderate at 100 µg/ml and profound at 1000 µg/ml. These results are significant as they suggest that NM has therapeutic potential in treatment of ovarian and cervical cancer.

 
 

A Novel Nutrient Mixture Induces Apoptosis in Human Prostate Cancer Cell Line Du-145

M.W. Roomi, N.W. Roomi, V. Ivanov, A. Niedzwiecki and M. Rath
Dr. Rath Research Institute, 1260 Memorex Drive, Santa Clara, CA 95050

Presented at: 
Apoptosis in Drug Discovery; San Diego, CA; March 22-23, 2007

Published in: 
Proccedings of the GTCbio Conference on Apoptosis in Drug Discovery

Abstract

Introduction:
Prostate cancer, the major cancer affecting males and second most deadly cancer in the United States, primarily affects males aged 55 and older and African Americans more than Caucasians. Once prostate cancer has metastasized, current treatment methods are generally ineffective. We have identified a novel antineoplastic agent, a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract that demonstrates a broad spectrum of antitumor activity against a number of human cancer cell lines.

Objective:
We investigated whether the cytotoxic effect of NM on DU145 was consistent with its apoptotic effect on these cells.

Method and Materials: 
Prostate cancer DU-145 (ATCC) cells were cultured in MEM and treated with NM in different concentrations, 0, 100, 500 and 1000 µg/ml concentration. Growth inhibition was detected by MTT assay, morphology by H&E staining, and apoptosis by Live Green Caspases Detection Kit.

Results:
NM was slightly toxic to DU145 cells at 100-µg/ml and profound at 500 and 1000 µg/ml. DU-145 cells treated with NM at 500 and 1000 µg/ml concentration and stained by H&E demonstrated obvious apoptotic cells. Apoptotic cells showed shrinkage with darkly stained and condensed nuclei and strongly acidophilic cytoplasm. Using caspases kit, slight apoptosis was observed at 100 µg, moderate at 500 and significant at 1000 µg/ml concentrations.

Conclusions:
The results suggest that NM induces apoptosis in prostate cancer cell line DU-145 and raises the possibility of its use as a new chemotherapeutic a gent.

Comment

A novel antineoplastic agent, a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract demonstrates a broad spectrum of antitumor activity against a number of human cancer cell lines. We investigated whether the cytotoxic effect of NM on DU145 was consistent with its apoptotic effect on these cells. DU-145 cells treated with NM at 500 and 1000 µg/ml demonstrated profound toxicity and obvious apoptotic cells morphologically. Using caspases kit, slight apoptosis was observed at 100 µg, moderate at 500 and significant at 1000 µg/ml concentrations. The results are significant as they suggest NM as a therapeutic agent for prostate cancer, the major cancer affecting males and second most deadly cancer in the United States.

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