ABSTRACT
The influence of cytostatically active alkyllysophospholipid analogs with different chemical structure on IP3 (inositol-1,4,5-trisphosphate) formation and intracellular Ca++ concentration was studied in human mammary epithelial cells before and after transfection with v-erb B oncogene DNA. Transformed cells showed an increased IP3 formation compared with normal cells. In the presence of ALP (alkyllysophospholipid) analogs IP3 formation is inhibited more strongly in transformed cells than in normal cells, dependent on the structure of ALP analogs. Furthermore, these compounds increased [Ca++]i (intracellular Ca++ concentration) in transformed cells much more strongly than in normal cells. From these results, obtained in pursuit of an oncogene-related cancer treatment, it follows that ALP analogs may inhibit processes in signal transduction in cells more strongly after transfection with a defined oncogene than before.
Subject(s)
Breast/drug effects , Inositol 1,4,5-Trisphosphate/biosynthesis , Lysophospholipids/pharmacology , Oncogene Proteins v-erbB/physiology , Phospholipid Ethers/pharmacology , Alpharetrovirus/genetics , Alpharetrovirus/physiology , Breast/cytology , Breast/metabolism , Cell Line, Transformed , Cell Transformation, Viral , Cells, Cultured , DNA, Viral/genetics , Dose-Response Relationship, Drug , Humans , Oncogene Proteins v-erbB/genetics , Signal Transduction/drug effects , TransfectionABSTRACT
Previous investigations in human precancerous and cancerous tissues identified subsets of cells that were different in respect to a heparin-induced increase in fluorescence intensity (IFI) monitored by flow cytometry. We suggested that differences in IFI were due to different chromatin types in the cells and related to different transcriptional capacities. This study is to prove our suggestion. Heparin-induced IFI was measured in the human breast cell line H184A1N4 cultured under different, defined growth conditions. Cells in quiescence obtained from a culture deprived of serum and other supplements essential for growth or from the confluent state revealed a higher IFI than cells restimulated to proliferation by addition of complete medium; here a reduced IFI was found. Changes in the magnitude of heparin-induced IFI precede changes in cell-cycle stage distribution by at least 6.5 h. We conclude that the heparin-induced effects, as revealed by flow cytometry, reflect changes in the accessibility of chromatin to transcription in different stages of proliferative activity. The results confirm our conclusions from previous findings with clinical material.
