INDUCTION OF APOPTOTIC PATHWAY IN CANCER CELL LINES

Bethune-Cookman College, Daytona Beach FL

Investigators

Linked publications & trials

Abstract

Growth of tissue in both normal and disease depends on the balance between cellular proliferation and programmed cell death (apoptosis). Decision to cell hormones, calcium balance, ATP, p53, Bcl-2 family and others. Calcium has been known to be a key regulator of cellular proliferation. Previously, it has been shown that high calcium concentration (20-50 mM) completely degrades important proteins like Hsp70, calcineurin, calmodulin, ubiquitin, casein and m-calpain in vitro (Sen 1996). The ability of calcium to degrade proteins depends on PESTDY sequence. For this reason, it could be hypothesized that calcium in combination with anti-angiogenic agents, electron transport chain inhibitors, and neoplastic agents would induce apoptosis more efficiently at a lower concentration than without calcium. Specific biochemical markers for apoptosis such as DNA fragmentation, cytochrome c release, CPP32/caspase-3, PARP, Bcl-2, Bax and p53 expression have been selected to measure the effectiveness of chemical agents to induce apoptosis in cultured human breast cancer cells (MCF-7 & BT-20). The cells will be exposed to various concentrations of calcium chloride (1-5 nM) in combination with rotenone (10nM-10muM), thalidomide (10nM- 10muM), methotrexate (10nM-10muM), 2-methoxyestradiol (10 nM- 10muM) and buthionine sulfoximine (10 nM-10muM) to see how effectively it induces apoptosis. Since calcium is one of the triggers for apoptosis and when combined with other apoptotic agents will show greater additive effect at a much lower dose than when treated with a single agent or in combination with other agents without calcium. Exploitation of cell's inherent apoptotic apoptotic mechanism would be a better therapeutic alternative for cancer. This study will provide a better understanding of calcium's potential as a therapeutic agent in the war against cancer.