title: Apoptosis of human breast carcinoma cells in the presence of cis-platin and L-/D-PPMP: IV. Modulation of replication complexes and glycolipid: Glycosyltransferases.
abstract: Apoptosis of human breast carcinoma cells (SKBR-3, MCF-7, and MDA-468) has been observed after treatment of these cells with anti-cancer drug cis-platin and glycosphingolipid biosynthesis inhibitor L- and D-PPMP, respectively. These drugs initiated apoptosis in a dose-dependent manner as measured by phenotypic morphological changes, by binding of a fluorescent phophatidyl serine-specific dye (PSS-380) onto the outer leaflet of the cell membranes, and by activation of caspases, -3, -8, and -9. It was observed that in two hours very little apoptotic process had started but predominant biochemical changes occurred after 6 h. DNA degradation started after 24 hours of drug treatment. However, very little is known about the stability of the ';Replication Complexes'' during the apoptotic process. DNA helicases are motor proteins that catalyze the melting of genomic DNA during its replication, repair, and recombination processes. Previously, DNA helicase-III was characterized as a component of the replication complexes isolated from embryonic chicken brains as well as breast and colon carcinoma cells. Helicase activities were measured by a novel method (ROME assay), and DNA polymerase-alpha activities were determined by regular chain extension of the nicked ACT-DNA, by determining values obtained from +/- aphidicolin-treated incubation mixtures. In all three breast carcinoma cell lines, a common trend was observed: a decrease of activities of DNA polymerase-alpha and Helicase III. A sharp decrease of activities of the glycolipid sialyltransferases: SAT-2 (CMP-NeuAc; GD3 alpha2-8 sialyltransferase) and SAT-4 (CMP-NeuAc: GM1a alpha2-3 sialyltransferase) was observed in the apoptotic carcinoma cells treated with L-PPMP compared with cis-platin.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Boyle
authors.forename: Rui
authors.initials: R
authors.lastname: Ma
authors.forename: Narendra
authors.initials: N
authors.lastname: Tuteja
authors.forename: Sipra
authors.initials: S
authors.lastname: Banerjee
authors.forename: Subhash
authors.initials: S
authors.lastname: Basu
mesh_terms: D000123:Acetyltransferases / Q000187:drug effects / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology*; D002240:Carbohydrate Sequence; D020169:Caspases / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D004265:DNA Helicases / Q000187:drug effects / Q000378:metabolism; D004256:DNA Polymerase I / Q000187:drug effects / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006017:Glycolipids / Q000378:metabolism*; D016695:Glycosyltransferases / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009025:Morpholines / Q000494:pharmacology*; D010718:Phosphatidylserines / Q000378:metabolism; D012799:Sialyltransferases / Q000187:drug effects / Q000378:metabolism; D013107:Sphingolipids / Q000494:pharmacology*; D000099228:Diamine N-Acetyltransferase
chemical_list: D000123:Acetyltransferases; D000970:Antineoplastic Agents; D020169:Caspases; D002945:Cisplatin; D004265:DNA Helicases; D004256:DNA Polymerase I; D006017:Glycolipids; D016695:Glycosyltransferases; D009025:Morpholines; D010718:Phosphatidylserines; D012799:Sialyltransferases; D013107:Sphingolipids; D000099228:Diamine N-Acetyltransferase; C095823:1-phenyl-2-palmitoylamino-3-morpholino-1-propanol; C019916:CMP-N-acetylneuraminic acid-monosialoganglioside sialyltransferase