Roomi MW, Bhanap B, Niedzwiecki A, Rath M
Apoptosis (Open Access e-book) ISBN: 978-93-86337-72-6; Publisher: Avid Science In Press: 2018 (http://www.avidscience.com/book/apoptosis/)
Renal cell carcinoma (RCC) is highly erratic and unpredictable even when diagnosed at an early stage. The incidence of RCC is higher among African Americans than among Caucasians. The male to female ratio is 2:1. Except for blood in the urine, other symptoms such as abdominal, back or flank pain, weight loss, and abnormal blood counts, are vague signs and symptoms that delay the diagnosis.
Therefore, at the time of the diagnosis, the RCC is already metastasized reducing the five-year-survival to 60% in RCC patients.A novel nutrient mixture (NM) containing green tea extract, ascorbic acid, lysine, and proline exhibited anticancer effects in various cancers. In this study, we investigated whether the anti-cancer properties of NM were due to the induction of apoptosis. The RCC cells (786-0) were cultured in complete DME media and the cells were treated with NM at 0-1000 µg/ml concentration. Cell cytotoxicity was measured by MTT assay, u-PA, MMP and TIMPs were measured by gelatinase, fibrin, and reverse zymography, cell morphology by H&E staining, apoptosis by Live Green Caspases. NM showed no significant effect on RCC cell growth, exhibiting slight toxicity at 100 µg and significant at 500 and 1000 µg/ml. RCC 786-0 cells treated at 500 and 1000 µg/ml NM and stained by H&E demonstrated obvious apoptotic cells with shrinkage and darkly stained and condensed nuclei and strong acidophilic cytoplasm. Using Live Green Caspases kit a significant number of early and late apoptotic RCC cells were seen at 500 and 1000 µg/ml NM. Our results suggest that NM induces apoptosis, which in turn down regulated u-PA and MMP and up regulated TIMPs. Therefore, NM may be a new therapeutic strategy for kidney cancer patients and deserves further investigation as a potential agent in the treatment of this malignancy.
Keywords: Kidney Cancer; 786-0; Apoptosis; Cytotoxicity; MTT; Live Green Caspase