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: Patrick J Boyle, Rui Ma, Narendra Tuteja, Sipra Banerjee, Subhash Basu
• Journal: Glycoconjugate journal (Glycoconj J) Vol. 23 Issue 3-4 Pg. 175-87 (May 2006) ISSN: 0282-0080 [Print] United States
• PMID: 16691501 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol
• Antineoplastic Agents
• Glycolipids
• Morpholines
• Phosphatidylserines
• Sphingolipids
• Acetyltransferases
• diamine N-acetyltransferase
• Glycosyltransferases
• Sialyltransferases
• CMP-N-acetylneuraminic acid-monosialoganglioside sialyltransferase
• DNA Polymerase I
• Caspases
• DNA Helicases
• Cisplatin

Topics

• Acetyltransferases (drug effects, metabolism)
• Antineoplastic Agents (pharmacology)
• Apoptosis
• Breast Neoplasms (drug therapy, pathology)
• Carbohydrate Sequence
• Caspases (drug effects, metabolism)
• Cell Line, Tumor
• Cisplatin (pharmacology)
• DNA Helicases (drug effects, metabolism)
• DNA Polymerase I (drug effects, metabolism)
• DNA Replication (drug effects, physiology)
• Dose-Response Relationship, Drug
• Female
• Glycolipids (metabolism)
• Glycosyltransferases (drug effects, metabolism)
• Humans
• Molecular Sequence Data
• Morpholines (pharmacology)
• Phosphatidylserines (metabolism)
• Sialyltransferases (drug effects, metabolism)
• Sphingolipids (pharmacology)