A nutrient mixture modulates ovarian ES-2 cancer progression by inhibiting xenograft tumor growth and cellular MMP secretion, migration and invasion

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 3 × 106 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 × length × 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.