title: Synergistic action of cisplatin and echistatin in MDA-MB-231 breast cancer cells.
abstract: The aim of our study was to determine whether the use of cisplatin in the presence echistatin in MDA-MB-231 breast cancer cells leads to a reduction of toxic effects associated with the use of cisplatin. The expression of β1-integrin and insulin-like growth factor 1 receptor (IGF-IR), signaling pathway protein expression: protein kinase B (AKT), mitogen-activated protein kinases (ERK1/ERK2), nuclear factor kappa B (NFκB), and caspase-3 and -9 activity was measured after 24 h of incubation with tested compounds to explain detailed molecular mechanism of induction of apoptosis. The viability of MDA-MB-231 breast cancer cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Annexin V-FITC/propidium iodide staining assay was performed to detect the induction of apoptosis. Inhibition DNA biosynthesis was determined by [3H]thymidine incorporation into DNA. The expression of of β1-integrin, IGF-IR, AKT, ERK1/ERK2, NFκB, caspase-3 and -9 was evaluated using Western blot. The results suggest that treatment of MDA-MB-231 breast cancer cells for 24 h cisplatin plus echistatin severely inhibits cell growth and activates apoptosis by upregulation of caspase-3 and -9 expressions. The effect was stronger than treatment cisplatin and echistatin alone. In this study, we have found that cisplatin plus echistatin treatment decreases collagen biosynthesis in MDA-MB-231 breast cancer cells stronger than the individual compounds. The inhibition was found to be dependent on the β1-integrin and IGF receptor activation. A significant reduction of ERK1/ERK2, AKT expression in cancer cells after cisplatin plus echistatin treatment was also found. The cancer cells treated by echistatin, cisplatin, and in particular the combination of both compounds drastically increased expression of NFκB transcription factor. Our results suggest that combined therapy cisplatin plus echistatin is a possible way to improve selectiveness of cisplatin. This mechanism probably is due to downregulation of expression of β1-integrin and IGF-IR receptors, and the signaling pathway proteins induced by these receptors. Our results suggest that therapy cisplatin plus echistatin is a possible way to improve selectiveness of cisplatin.
authors.affiliation: Department of Synthesis and Technology of Drugs, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland. robert.czarnomysy@umb.edu.pl.
authors.forename: Robert
authors.identifier: http://orcid.org/0000-0002-7148-6111
authors.initials: R
authors.lastname: Czarnomysy
authors.affiliation: Department of Medicinal Chemistry, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Arkadiusz
authors.initials: A
authors.lastname: Surażyński
authors.affiliation: Department of Biotechnology, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Bożena
authors.initials: B
authors.lastname: Popławska
authors.affiliation: Department of Medicinal Chemistry, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Edyta
authors.initials: E
authors.lastname: Rysiak
authors.affiliation: Department of Synthesis and Technology of Drugs, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Pawłowska
authors.affiliation: Department of Biotechnology, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Czajkowska
authors.affiliation: Department of Synthesis and Technology of Drugs, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Bielawski
authors.affiliation: Department of Biotechnology, Medical University of Bialystok, Kilinskiego 1, 15-089, Bialystok, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Bielawska
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000819:agonists / Q000494:pharmacology*; D004357:Drug Synergism; D005260:Female; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins / Q000378:metabolism; D010455:Peptides / Q000819:agonists / Q000494:pharmacology*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D010455:Peptides; C057986:echistatin; D002945:Cisplatin
keywords: Apoptosis; Breast cancer cells; Cell signaling; Cisplatin; Echistatin