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

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

Presented at: 55th Annual Meeting of ToxExpo, March 13-17, 2016, New Orleans, LA

Published in: The Toxicologist 150 (1), 2016 Abstract #2023, pg 240


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. We investigated the effect of a nutrient mixture (EPQ) containing ascorbic acid, lysine, proline, quercetin and green tea extract in vivo and in vitro on human ovarian cancer ES-2 cell line. In vivo, athymic female nude mice (n=12) were inoculated with 3x106 ES-2 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% EPQ. Four weeks later, the mice were sacrificed and their tumors were excised, weighed 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 EPQ in vitro on ES-2 cells, measuring cell proliferation by MTT assay, MMP secretion by zymography, invasion through Matrigel, migration by scratch test and morphology by H&E staining. EPQ inhibited tumor weight and burden of ES-2 tumors by 59.2% (p <0.0001) and 59.7% (p<0.0001), respectively. In vitro, EPQ exhibited 35% (p<0.0001) toxicity over the control at 1000 µg/ml concentration. Zymography demonstrated only MMP-2 with and without PMA, which was inhibited by EPQ in a dose dependent fashion, with near total inhibition at 1000 µg/ml. Migration by scratch test and Invasion through Matrigel were inhibited in a dose dependent manner with total block of invasion and migration at 500 µg/ml. These results suggest that EPQ has therapeutic potential in treatment of ovarian cancer.

Current treatment methods for ovarian cancer are generally ineffective and have not improved in 40 years.  Thus, there is a need for development of effective therapeutic agents for these cancers with minimal toxicity. Our studies demonstrated that the mixture of the non-toxic components of EPQ significantly inhibited the growth and tumor burden of ovarian cancer cell line ES-2 in vivo. In addition, invasive parameters, such as ES-2 cell line MMP-2 secretion, migration and invasion were significantly inhibited by EPQ in vitro. These findings are significant since they suggest potential of EPQ in treatment of ovarian cancer.