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01-01-2012 | Original Article

The marine sponge toxin agelasine B increases the intracellular Ca²⁺ concentration and induces apoptosis in human breast cancer cells (MCF-7)

Authors: Adriana A. Pimentel, Pimali Felibertt, Felipe Sojo, Laura Colman, Adriana Mayora, May Li Silva, Hector Rojas, Reinaldo Dipolo, Alírica I. Suarez, Reinaldo S. Compagnone, Francisco Arvelo, Ivan Galindo-Castro, Juan B. De Sanctis, Perla Chirino, Gustavo Benaim

Published in: Cancer Chemotherapy and Pharmacology | Issue 1/2012

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 Ca²⁺ concentrations were assessed with FURA 2 and by confocal microscopy. Determination of Ca²⁺-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 (IC₅₀ for MCF-7 = 2.99 μM; SKBr3: IC₅₀ = 3.22 μM vs. fibroblasts: IC₅₀ = 32.91 μM), while the IC₅₀ for PC-3 IC₅₀ = 6.86 μM. Agelasine B induced a large increase in the intracellular Ca²⁺ 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 Ca²⁺ from the endoplasmic reticulum (ER). We also demonstrated that agelasine B produces a potent inhibition of the ER Ca²⁺-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 [Ca²⁺]_i after blocking the SERCA activity. The reproduction of the effects of agelasine B on cell viability and on the [Ca²⁺]_I obtained on SKBr3 and PC-3 cancer cells strongly suggests the generality of the mechanism of action of this toxin.

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Title: The marine sponge toxin agelasine B increases the intracellular Ca2+ concentration and induces apoptosis in human breast cancer cells (MCF-7)
Authors: Adriana A. Pimentel
Pimali Felibertt
Felipe Sojo
Laura Colman
Adriana Mayora
May Li Silva
Hector Rojas
Reinaldo Dipolo
Alírica I. Suarez
Reinaldo S. Compagnone
Francisco Arvelo
Ivan Galindo-Castro
Juan B. De Sanctis
Perla Chirino
Gustavo Benaim
Publication date: 01-01-2012
Publisher: Springer-Verlag
Published in: Cancer Chemotherapy and Pharmacology / Issue 1/2012
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-011-1677-x

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