Retinoblastoma is one of the most common ocular malignancies in children under the age of six. Occasionally, retinoblastoma metastasizes to extraocular organs including the bone, lung and brain. Left untreated, retinoblastoma is fatal. At present, there is no effective treatment for metastatic retinoblastoma. We investigated the antineoplastic activity of a nutrient mixture (NM) (lysine, proline, ascorbic acid and green tea extract) at concentrations of 10, 50, 100, 500 and 1,000 μg/ml in triplicate at each dose in the human malignant retinoblastoma Y-79 cell line. The parameters used were cell proliferation, expression of matrix metalloproteinases (MMPs), invasion through Matrigel, morphology and apoptosis. Cell viability was assessed by trypan blue dye exclusion test. Invasion was evaluated through Matrigel and MMP activity by gelatinase zymography. H&E staining for morphological cell alterations and apoptotic studies using the Live Green Poly Caspase Detection kit were also conducted. The nutrient mixture at 10–100 μg/ml demonstrated approximately 25% toxicity towards Y-79 retinoblastoma cells and significant toxicity at 500 and 1,000 μg/ml. The Y-79 cells secreted only MMP-2 as demonstrated by zymography; the nutrient mixture had no effect on MMP-2 expression up to 100 μg/ml, but completely blocked it at 500 μg/ml. Importantly, Y-79 retinoblastoma cells were not invasive through Matrigel. H&E staining showed cell morphological changes related to apoptosis, which was confirmed using the Live Green Poly Caspase Detection kit. Our results suggest that this nutrient mixture, which inhibited cell proliferation, expression of MMP-2 and induced apoptosis, may be a candidate for further exploration for its therapeutic potential in metastatic retinoblastoma.

The prognosis of hepatocellular carcinoma (HCC) remains dismal despite any treatment. Matrix metalloproteinases (MMPs) have been researched for their role in tumor invasion and metastasis. Various cytokines, mitogens, growth factors, inducers, and inhibitors control MMP activities. In this article, we investigated the roles of these in the regulation of MMP-2, -9 secretions in HCC. Human HCC SK-Hep-1 was grown in appropriate media. At near confluence, the cells were washed with phosphate-buffered saline and incubated in serum-free media with PMA; TNF-α, IL-1β; lipopolysaccharide; epigallocatechin gallate (EGCG) and doxycycline (Dox) at various doses with and without PMA; a nutrient mixture (NM) containing lysine, proline, ascorbic acid, and EGCG with and without PMA at; and actinomycin D and cycloheximide at different doses. After 24 hours, the media were removed and analyzed. SK-Hep-1 expressed bands corresponding to MMP-2 and MMP-9. TNF-α showed an insignificant effect on MMP-2 at doses below 25 at which dose MMP-2 was virtually blocked and a moderate dose-dependent effect on MMP-9. Interleukin-1β demonstrated an insignificant effect on MMP-2 at doses below 25 at which dose MMP-2 was completely blocked and enhanced effects on MMP-9. Lipopolysaccharide showed dose-dependent inhibition of MMP-2 and MMP-9. EGCG, Dox, and NM, without and with PMA, downregulated the expression of MMP-2 and MMP-9. Actinomycin D and cycloheximide also had dose-dependent inhibitory effects on MMPs. Our results showed that cytokines, mitogens, and inhibitors modulated SK-Hep-1 MMP-2 and MMP-9 secretion, suggesting the clinical use of especially potent and nontoxic MMP inhibitor as the NM in management of HCC.

Background: Tumor metastasis is a major cause for most cancer-related deaths. Melanoma is a serious cancer that metastasizes to other areas of the body, including the lungs, liver, brain, bones, or lymph nodes. Currently used cancer therapies are ineffective with a high degree of toxicity and patient mortality. Thus, any successful treatment for melanoma must target metastasis. Methods: We studied the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline, green tea extract, quercetin, and others, on the inhibition of melanoma growth and metastasis after inoculation of B16FO melanoma cells into the left kidney of female nude mice. Female athymic mice (n = 10) 8 to 10 weeks of age, were inoculated superficially in the left kidney with 5 × 10 5 B16FO melanoma cells in 100 µL of media. The right kidney was left untreated. After inoculation, the mice were randomly divided into 2 groups. The control group (n = 5) was fed a regular rodent chow diet, and the test group was given the same diet supplemented with 0.5% NM. The animals in control and the test groups were sacrificed 2 weeks later. Each animal’s abdominal cavity was opened, and the kidneys, lungs, liver, and spleen were excised and examined for tumor growth and metastasis. Results: The kidneys in the control group weighed 25% to 30% more than those in the NM group due to colonization of B16FO melanoma cells. No metastasis to the liver or spleen was observed in either of the groups. However, severe lung metastasis was observed in the control group and mild to moderate metastasis was observed in the NM group. Conclusion: These results show that the NM is effective in mitigating the growth of tumors in the kidney and metastases to the lung.

A majority of oral cancers is squamous cell carcinoma and tongue carcinomas comprise 30% of all oral cancers. Phytochemicals, herbal and dietary antioxidants have been reported to prevent cancers. A nutrient mixture containing, ascorbic acid, lysine, proline and green tea extract, among other nutrients, has previously been shown to exhibit a broad spectrum chemo preventative and therapeutic anti-cancer properties in a number of cell lines. In a recent study, we found that the nutrient mixture significantly inhibited acetaminophen induced hepatic and renal toxicity, and it suppressed carbon tetrachloride induced hepatic toxicity in ICR mice as well. This study was undertaken to determine if the nutrient mixture is useful in inhibiting various parameters of cancer progression on human tongue cancer cell line SC-255. SC-255 cells were grown in a Dulbecco's Eagle medium and treated with the nutrient mixture at 0, 10, 50, 100, 500 and 1000 µg/ml, in triplicate. The nutrient mixture exhibited 20% and 30% toxicity at 500 and 1000 µg/ml, respectively. Zymography demonstrated the expression of MMP-2 and MMP-9; and PMA treatment further enhanced MMP-9 activity. The nutrient mixture inhibited the secretion of MMP-2 and MMP-9 in a dose-dependent fashion with a total inhibition at 500 µg/ml. Matrigel invasion was significantly reduced by 40%, 80% and 100% at 100, 500 and 1000 µg/ml, respectively. The nutrient mixture also inhibited cell migration and induced apoptosis in a dose response fashion. Thus, the nutrient mixture may have a potential in the tongue cancer treatment.

Background: Cancer is the second leading cause of death in the world. Metastasis is the process by which tumor cells leave the primary site and form colonies in different locations. The risk of metastasis makes cancer difficult to treat and accounts for a 90 percent mortality rate. A major problem in studying metastasis has been a lack of suitable models that faithfully represent the metastatic process as it occurs in vivo. Purpose: In order to mimic the potential of tumor metastasis in humans, we used a highly aggressive B16FO murine melanoma cell line, and showed that B16FO melanoma can metastasize to various sites. In the current study we investigated whether B16FO cells inoculated in the non-typical organs in male nude mice and female C57BL6 mice have the potential to metastasize to distant organs. As such, we injected B16FO cells in prostate, seminal vesicles and the pancreas of male nude mice and in mammary pads of female C57BL6 mice. The animals were kept on recommended diet and sacrificed after three weeks. Results: We observed that while the animals did develop tumors at the sites of inoculation, the B16FO cells did not induce metastasis in any of the vital organs. It is possible that although B16FO are very aggressive cells, the three weeks were not enough for metastasis to develop. Otherwise it could be the tumor dormancy period. Conclusion: We conclude that out of the four inoculation sites we tested in this series of experiments such as the prostate, seminal vesicles, the pancreas and mammary pads of nude or C57BL6 mice are not the best route for future investigation of melanoma metastasis.