title: Anticancer Activity of New Synthetic α-Methylene-δ-Lactones on Two Breast Cancer Cell Lines.
abstract: Natural products are important leads in drug discovery. The search for effective plant-derived anticancer agents or their synthetic analogues has continued to be of interest to biologists and chemists for a long time. In this report, cytotoxicity and anticancer activity of new synthetic α-methylene-δ-lactones was tested against two breast cancer cell lines, invasive, hormone-independent MDA-MB-231 and hormone-dependent MCF-7. Cytotoxicity was examined using MTT assay. The ability to induce apoptosis and changes in mitochondrial membrane potential was studied by flow cytometry. The expression levels of pro- and anti-apoptotic genes were determined by quantitative real-time PCR. Cancer cell migration and invasion were assessed by wound healing and Matrigel assays. Additionally, secretion of proteins associated with invasiveness, metalloproteinase-9 (MMP-9) and urokinase plasminogen activator (uPA) was investigated using commercial ELISA kits and MMP-9 activity by gelatin zymography. A natural sesquiterpene lactone, parthenolide, was used as a positive control. Screening results showed all four analogues to be highly cytotoxic. The most potent compound of the series, 1-isopropyl-2-methylene-1,2-dihydrobenzochromen-3-one, designated DL-3, which reduced the number of viable MDA-MB-231 and MCF-7 cells with the IC50 values of 5.3 μM and 3.54 μM, respectively, was selected for further research. DL-3 activated the intrinsic pathway of apoptosis, associated with the loss of mitochondrial membrane potential and changes in Bax/Bcl-2 ratio. DL-3 also inhibited the movement of both types of breast cancer cells. Suppression of cell migration and invasion was the result of the decreased secretion of enzymes responsible for the degradation of the extracellular matrix, MMP-9 and uPA. These findings show that the synthetic α-methylene-δ-lactone, DL-3, displays potential to be further explored in the development of new anticancer agents.
authors.affiliation: Department of Biomolecular, Chemistry Medical University of Lodz, Lodz, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Wyrębska
authors.affiliation: Department of Biomolecular, Chemistry Medical University of Lodz, Lodz, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Gach
authors.affiliation: Department of Enzymology, Medical University of Lodz, Lodz, Poland.
authors.forename: Urszula
authors.initials: U
authors.lastname: Lewandowska
authors.affiliation: Department of Enzymology, Medical University of Lodz, Lodz, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Szewczyk
authors.affiliation: Department of Enzymology, Medical University of Lodz, Lodz, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Hrabec
authors.affiliation: Institute of Organic Chemistry, Technical University of Lodz, Lodz, Poland.
authors.forename: Jakub
authors.initials: J
authors.lastname: Modranka
authors.affiliation: Institute of Organic Chemistry, Technical University of Lodz, Lodz, Poland.
authors.forename: Rafał
authors.initials: R
authors.lastname: Jakubowski
authors.affiliation: Institute of Organic Chemistry, Technical University of Lodz, Lodz, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Janecki
authors.affiliation: Central Laboratory, Medical University of Lodz, Lodz, Poland.
authors.forename: Jacek
authors.initials: J
authors.lastname: Szymański
authors.affiliation: Department of Biomolecular, Chemistry Medical University of Lodz, Lodz, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Janecka
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000188:drug therapy*; D045744:Cell Line, Tumor / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D003374:Coumarins / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D006801:Humans; D061986:MCF-7 Cells / Q000187:drug effects*; D020780:Matrix Metalloproteinase 9 / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D014568:Urokinase-Type Plasminogen Activator / Q000187:drug effects / Q000378:metabolism
chemical_list: C000596360:1-isopropyl-2-methylene-1,2-dihydrobenzochromen-3-one; D000970:Antineoplastic Agents; D003374:Coumarins; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9