Mitochondria-mediated apoptosis operating irrespective of multidrug resistance in breast cancer cells by the anticancer agent prodigiosin.

Prodigiosin (PG) is a red pigment produced by Serratia marcescens with pro-apoptotic activity in haematopoietic and gastrointestinal cancer cell lines, but no marked toxicity in non-malignant cells. Breast cancer is the most frequent malignancy among women in the European Union and better therapies are needed, especially for metastatic tumors. Moreover, multidrug resistance is a common phenomenon that appears during chemotherapy, necessitating more aggressive treatment as prognosis worsens. In this work, we extend our experiments on PG-induced apoptosis to breast cancer cells. PG was potently cytotoxic in both estrogen receptor positive (MCF-7) and negative (MDA-MB-231) breast cancer cell lines. Cytochrome c release, activation of caspases-9, -8 and -7 and cleavage of poly (ADP-ribose) polymerase protein typified the apoptotic event and caspase inhibition revealed that PG acts via the mitochondrial pathway. In a multidrug-resistant subline of MCF-7 cells that over-expresses the breast cancer resistance protein, the cytotoxic activity of PG was slightly reduced. However, flow-cytometry analysis of PG accumulation and efflux in MCF-7 sublines showed that PG is not a substrate for this resistance protein. These results suggest that PG is an interesting and potent new pro-apoptotic agent for the treatment of breast cancer even when multidrug resistance transporter molecules are present.

[1]  G. Loo,et al.  Disruption of mitochondria during tocotrienol-induced apoptosis in MDA-MB-231 human breast cancer cells. , 2004, Biochemical pharmacology.

[2]  M. Péč,et al.  Induction of apoptosis in estrogen dependent and independent breast cancer cells by the marine terpenoid dehydrothyrsiferol. , 2003, Biochemical pharmacology.

[3]  R. Vossen,et al.  Complete sequencing of TP53 predicts poor response to systemic therapy of advanced breast cancer. , 2000, Cancer research.

[4]  S. B. Han,et al.  T-cell specific immunosuppression by prodigiosin isolated from Serratia marcescens. , 1998, International journal of immunopharmacology.

[5]  E. Slee,et al.  To die or not to die: how does p53 decide? , 2004, Oncogene.

[6]  A. Thor,et al.  Reconstitution of caspase 3 sensitizes MCF-7 breast cancer cells to doxorubicin- and etoposide-induced apoptosis. , 2001, Cancer research.

[7]  J. Schellens,et al.  Breast cancer resistance protein is localized at the plasma membrane in mitoxantrone- and topotecan-resistant cell lines. , 2000, Cancer research.

[8]  D. Green.  Apoptotic Pathways The Roads to Ruin , 1998, Cell.

[9]  A. Duncan,et al.  Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. , 1992, Science.

[10]  H. Kamata,et al.  A possible immunosuppressant, cycloprodigiosin hydrochloride, obtained from Pseudoalteromonas denitrificans. , 1997, Biochemical and biophysical research communications.

[11]  D. Housman,et al.  p53-dependent apoptosis modulates the cytotoxicity of anticancer agents , 1993, Cell.

[12]  R. Pérez-Tomás,et al.  Activation of protein kinase C for protection of cells against apoptosis induced by the immunosuppressor prodigiosin. , 2002, Biochemical pharmacology.

[13]  Alan G. Porter,et al.  Caspase-3 Is Required for DNA Fragmentation and Morphological Changes Associated with Apoptosis* , 1998, The Journal of Biological Chemistry.

[14]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[15]  T. Litman,et al.  Molecular cloning of cDNAs which are highly overexpressed in mitoxantrone-resistant cells: demonstration of homology to ABC transport genes. , 1999, Cancer research.

[16]  P. Borst,et al.  Mammalian ABC transporters in health and disease. , 2002, Annual review of biochemistry.

[17]  H. Hirata,et al.  Cycloprodigiosin hydrochloride, a H+/Cl− symporter, induces apoptosis in human breast cancer cell lines , 2000, Journal of Cancer Research and Clinical Oncology.

[18]  R. Pérez-Tomás,et al.  Effects of the proapoptotic drug prodigiosin on cell cycle-related proteins in Jurkat T cells. , 2003, Histology and histopathology.

[19]  J. Schellens,et al.  Potent and specific inhibition of the breast cancer resistance protein multidrug transporter in vitro and in mouse intestine by a novel analogue of fumitremorgin C. , 2002, Molecular cancer therapeutics.

[20]  J. Holden,et al.  The anti-neoplastic and novel topoisomerase II-mediated cytotoxicity of neoamphimedine, a marine pyridoacridine. , 2003, Biochemical pharmacology.

[21]  G. Pons,et al.  Aspirin induces apoptosis through mitochondrial cytochrome c release , 2000, FEBS letters.

[22]  H. Hirata,et al.  Cycloprodigiosin hydrochloride, H+/CL– symporter, induces apoptosis and differentiation in HL‐60 cells , 2000, International journal of cancer.

[23]  A. Mantovani,et al.  Characterization of the new immunosuppressive drug undecylprodigiosin in human lymphocytes: retinoblastoma protein, cyclin-dependent kinase-2, and cyclin-dependent kinase-4 as molecular targets. , 1997, Journal of immunology.

[24]  John Calvin Reed,et al.  Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. , 2000, Cancer research.

[25]  D. Colomer,et al.  Prodigiosin induces apoptosis of B and T cells from B-cell chronic lymphocytic leukemia , 2003, Leukemia.

[26]  G. Salvesen,et al.  Caspases: Intracellular Signaling by Proteolysis , 1997, Cell.

[27]  J. Menéndez,et al.  Effects of gamma-linolenic acid and oleic acid on paclitaxel cytotoxicity in human breast cancer cells. , 2001, European journal of cancer.

[28]  S. Pal,et al.  Curcumin induces apoptosis in human breast cancer cells through p53‐dependent Bax induction , 2002, FEBS letters.

[29]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[30]  S. Srinivasula,et al.  Cytochrome c and dATP-Dependent Formation of Apaf-1/Caspase-9 Complex Initiates an Apoptotic Protease Cascade , 1997, Cell.

[31]  G. Hortobagyi,et al.  Chemotherapy of metastatic breast cancer: what to expect in 2001 and beyond. , 2001, The oncologist.

[32]  R. Pérez-Tomás,et al.  Prodigiosin induces cell death and morphological changes indicative of apoptosis in gastric cancer cell line HGT-1. , 2001, Histology and histopathology.

[33]  L. Doyle,et al.  A multidrug resistance transporter from human MCF-7 breast cancer cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[34]  R L Juliano,et al.  A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. , 1976, Biochimica et biophysica acta.

[35]  E. Giralt,et al.  Prodigiosin from the supernatant of Serratia marcescens induces apoptosis in haematopoietic cancer cell lines , 2000, British journal of pharmacology.

[36]  T. Druley,et al.  From MDR to MXR: new understanding of multidrug resistance systems, their properties and clinical significance , 2001, Cellular and Molecular Life Sciences CMLS.

[37]  Y. Jeon,et al.  Prodigiosin blocks T cell activation by inhibiting interleukin-2Ralpha expression and delays progression of autoimmune diabetes and collagen-induced arthritis. , 2001, The Journal of pharmacology and experimental therapeutics.

[38]  P. Gnocchi,et al.  Synthesis and immunosuppressive activity of novel prodigiosin derivatives. , 2000, Journal of medicinal chemistry.

[39]  J. Trent,et al.  Different mechanisms of decreased drug accumulation in doxorubicin and mitoxantrone resistant variants of the MCF7 human breast cancer cell line. , 1991, British Journal of Cancer.