New findings of minority benzo[c]phenenthridine alkaloids
|Original title:||New findings of minority benzo[c]phenenthridine alkaloids|
|Authors:||Iva Slaninová, Jiří Šinkora, Zdenka Slunská, Eva Táborská|
|Field:||Genetics and molecular biology|
|Type:||Article in Proceedings|
Quaternary benzo[c]phenanthridine alkaloids (QBA) are naturally occurring products with noteworthy antiproliferative activity and fluorescence characteristics. QBA are commonly isolated from plants of Fumariaceae, Papaveraceae, Ranunculaceae and Rutaceae families. Among them, sanguinarine, chelerythrine and fagaronin are the best recognized and characterized. We focused on minor QBA derivatives - macarpine (MA), sanguirubine (SR) and chelirubine (CHR) - and studied their antiproliferative, pro-apoptotic and anti-microtubule activity on human cell lines. In addition, we have observed interesting fluorescent characteristics of these molecules upon intercalating into DNA in live cells. We have proved that MA, SR, CHR can be used as supravital fluorescent DNA probes both in fluorescence microscopy and flow cytometry. When added to intact cell suspensions, water stock solutions of the alkaloids stain the cells upon brief (in seconds) incubation at low final concentration (0,01-0,001 mg/ml). We have also evaluated the convenience of minor QBA as supravital nucleic acid probes for time effective multiparameter immunophenotyping of peripheral blood and bone marrow leukocytes. Heparinized peripheral blood or bone marrow aspirate cells were stained on the surface with a combination of fluorochrome-conjuagted monoclonal antibodies directed against surface markers. Then, an appropriate amount of individual QBA was added and, upon dilution, the samples were acquired on a standard (FACSCalibur) or polychromatic (CyAn) flow cytometer. Our results show that spectral characteristics of individual QBA allow for simultaneous surface immunophenotyping and semiquantitative DNA determination. MA, SR and CHR are excitable by common argon lasers (488 nm) emmiting at the range 575 - 750 nm (i.e. fluorescence channels 2-5). Fluorescence channels 1 and 5 – 7 are available for surface immunophenotyping on FACSCalibur and CyAn, respectively.