title: Cytotoxicity and induction of apoptosis by formamidinodoxorubicins in comparison to doxorubicin in human ovarian adenocarcinoma cells.
abstract: OBJECTIVE
  In this study we investigated the effect of DOX and five of its derivatives containing a formamidine group (NCHNRR) at the 3' position with pyrrolidine (DOX-F PYR), piperidine (DOX-F PIP), morpholine (DOX-F MOR), N-methylpiperazine (DOX-F PAZ) and hexamethyleneimine (DOX-F HEX) ring on SKOV-3 ovarian cancer cells. We have focused on the anti-proliferative activity and the value of apoptosis induced by tested analogues.
  
  
  METHODS
  The following methods were used: spectrophotometric assay with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide); fluorimetric assays - double staining with Hoechst 33258 and propidium iodide (PI), measurement of caspase-3 activity; flow cytometry methods - phosphatidylserine (PS) externalization using Annexin V-FITC and PI fluorochromes, and TUNEL assay.
  
  
  RESULTS
  All of the investigated derivatives were considerably more cytotoxic to the SKOV-3 cell line than DOX. The predominant type of cell death induced by the anthracycline analogues was apoptosis. Necrotic cells represented only a small percentage (<5%) of all cells. The number of apoptotic cells was dependent on the compound and the incubation time. Moreover, a significant increase in caspase-3 activity, DNA fragmentation, and morphological changes in ovarian cells were observed predominantly in new DOX analogues.
  
  
  CONCLUSIONS
  All new formamidine derivatives of DOX were effective against ovarian cancer cells. They induced mainly the apoptotic pathway of cell death mediated by caspase-3. The most promising results were obtained for DOX-F MOR and DOX-F PAZ. The least potent was DOX-F HEX.
authors.affiliation: Department of Thermobiology, Institute of Biophysics, Faculty of Biology and Environmental Protection, Lodz University, Pomorska 141/143, 90-236 Lodz, Poland. Electronic address: aszwar@biol.uni.lodz.pl.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Marczak
authors.affiliation: Department of Thermobiology, Institute of Biophysics, Faculty of Biology and Environmental Protection, Lodz University, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Denel-Bobrowska
authors.affiliation: Department of Thermobiology, Institute of Biophysics, Faculty of Biology and Environmental Protection, Lodz University, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: Aneta
authors.initials: A
authors.lastname: Rogalska
authors.affiliation: Department of Modified Antibiotics, Institute of Biotechnology and Antibiotics, 5 Staroscinska St., 02-516 Warsaw, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Łukawska
authors.affiliation: Department of Modified Antibiotics, Institute of Biotechnology and Antibiotics, 5 Staroscinska St., 02-516 Warsaw, Poland.
authors.forename: Irena
authors.initials: I
authors.lastname: Oszczapowicz
mesh_terms: D000230:Adenocarcinoma; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D005260:Female; D006801:Humans; D009336:Necrosis / Q000139:chemically induced; D010051:Ovarian Neoplasms
chemical_list: D000903:Antibiotics, Antineoplastic; D004317:Doxorubicin; D053148:Caspase 3
keywords: Anthracycline derivatives; Apoptosis; Caspase-3; Cytotoxicity; Necrosis; TUNEL assay