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The marine sponge toxin agelasine B increases the intracellular Ca <sup>2+</sup> concentration and induces apoptosis in human breast cancer cells (MCF-7)

Abstract:

Purpose: In search for new drugs derived from natural products for the possible treatment of cancer, we studied the action of agelasine B, a compound purified from a marine sponge Agelas clathrodes. Methods: Agelasine B was purified from a marine sponge Agelas clathrodes and assayed for cytotoxicity by MTT on two human breast cancer cells (MCF-7 and SKBr3), on a prostate cancer cells (PC-3) and on human fibroblasts. Changes in the intracellular Ca 2+ concentrations were assessed with FURA 2 and by confocal microscopy. Determination of Ca 2+-ATPase activity was followed by Pi measurements. Changes in the mitochondria electrochemical potential was followed with Rhodamine 123. Apoptosis and DNA fragmentation were determined by TUNEL experiments. Results: Upon agelasine B treatment, cell viability of both human breast cancer cell lines was one order of magnitude lower as compared with fibroblasts (IC 50 for MCF-7 = 2.99 μM; SKBr3: IC 50 = 3.22 μM vs. fibroblasts: IC 50 = 32.91 μM), while the IC 50 for PC-3 IC 50 = 6.86 μM. Agelasine B induced a large increase in the intracellular Ca 2+ concentration in MCF-7, SKBr3, and PC-3 cells. By the use of confocal microscopy coupled to a perfusion system, we could observe that this toxin releases Ca 2+ from the endoplasmic reticulum (ER). We also demonstrated that agelasine B produces a potent inhibition of the ER Ca 2+-ATPase (SERCA), and that this compound induced the fragmentation of DNA. Accordingly, agelasine B reduced the expression of the anti-apoptotic protein Bcl-2 and was able to activate caspase 8, without affecting the activity of caspase 7. Conclusions: Agelasine B in MCF-7 cells induce the activation of apoptosis in response to a sustained increase in the [Ca 2+] i after blocking the SERCA activity. The reproduction of the effects of agelasine B on cell viability and on the [Ca 2+] I obtained on SKBr3 and PC-3 cancer cells strongly suggests the generality of the mechanism of action of this toxin. © 2011 Springer-Verlag.