Biological effects of selected benzo(c)phenanthridine alkaloids

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Authors

SLANINOVÁ Iva VLKOVÁ Marcela ŠINKORA Jiří TÁBORSKÁ Eva

Year of publication 2004
Type Article in Proceedings
Conference Sborník XII. Cytoskeletálního klubu
MU Faculty or unit

Faculty of Medicine

Citation
Field Morphological specializations and cytology
Keywords benzo(c)phenanthridine alkaloids; cytotoxicity; apoptosis; microtubule;
Description Quaternary benzo[c]phenanthridine alkaloids (QBA) are relatively small group of isoquinoline alkaloids interesting for their chemistry and biological activities. They are distributed in a number of plant species of Papaveraceae, Fumariaceae and Rutaceae families. Sanguinarine and chelerythrine are the two best known QBA and their biological activities have been widely studied (Walterová et al., 95). In addition to them about 10 other QBA differing only by substitution on the aromatic rings have been isolated as minor components in several species. The biological effects of these minor QBA have not been described yet. In this preliminary study the biological efect of sanguinarine (SA), chelerythrine (CHE) and their derivatives sanguirubine (SR), chelirubine (CHR) and macarpine (MA) were tested. The alkaloids were isolated from the species Macleaya microcarpa by the common procedure (Dostál et al., 1995). One normal (fibroblasts) and 3 tumor cell lines (HeLa; A431; HL-60) were used as a model object. Cytotoxicity (IC50), apoptosis and anti-microtubular effect on living cells were studied. After 72 hours, MTT assay was performed to evaluate the in vitro cytotoxicity induced by tested alkaloids at concentration 0,01-5 mg/ml. IC50 values for individual alkaloids were determined. There were not radical differences in the reaction of individual cell lines. Our preliminary results show that HL-60 and human fibroblasts were the most sensitive cell culture lines. Cytotoxicity (IC50; mg/ml) of individual alkaloids descended in this order SA (0,03-0,1)> CHE (0,03-0,3) > MA (0,03-0,3) > SR(0,05-3) > CHR(1-5). For apoptosis detection morphology of nuclei of alkaloid treated HL-60 and HeLa cells after DAPI staining was studied and flow cytometric method used Anexin V-FITC and PI (APOTESTTM-FITC, DAKO Cytomation) was applied to HL-60 cell line. SR, SA, MA and CHE appeared to be inductors of apoptosis. The effect on cytoskeleton was also studied. Actin was not seriously affected by any of the studied alkaloids, while microtubules were damaged by SR and MA.
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