SLANINOVÁ, Iva, Jiří SLANINA and Eva TÁBORSKÁ. Quaternary Benzo[c]phenanthridine Alkaloids - Novel Cell Permeant and Red Fluorescing DNA Probes. Cytometry Part A. UNITED STATES: John Wiley & Sons, Inc., 2007, 71A, No 9, p. 700-708. ISSN 1552-4922.
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Basic information
Original name Quaternary Benzo[c]phenanthridine Alkaloids - Novel Cell Permeant and Red Fluorescing DNA Probes
Name in Czech Kvartérní benzo[c]fenanthridinové alkaloidy - nové supravitální červeně fluoreskující DNA sondy
Authors SLANINOVÁ, Iva (203 Czech Republic, guarantor), Jiří SLANINA (203 Czech Republic) and Eva TÁBORSKÁ (203 Czech Republic).
Edition Cytometry Part A, UNITED STATES, John Wiley & Sons, Inc. 2007, 1552-4922.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10600 1.6 Biological sciences
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 2.978
RIV identification code RIV/00216224:14110/07:00020661
Organization unit Faculty of Medicine
UT WoS 000249187800011
Keywords in English benzophenanthridine alkaloids; cell-permeant DNA dye; chelirubine; flow cytometry; fluorescence microscopy; DNA content; cell cycle; macarpine; multicolor analysis; sanguinarine
Tags Benzophenanthridine Alkaloids, cell cycle, cell-permeant DNA dye, Chelirubine, DNA content, flow cytometry, fluorescence microscopy, Macarpine, multicolor analysis, sanguinarine
Tags International impact, Reviewed
Changed by Changed by: prof. MUDr. Iva Slaninová, Ph.D., učo 2105. Changed: 22/6/2009 12:04.
Background: Quaternary benzo[c]phenanthridine alkaloids (QBAs) are naturally occurring compounds isolated from plants of Fumariaceae, Papaveraceae, Ranunculaceae and Rutaceae families. In addition to wide biological activities, they are attractive for their fluorescence. We observed interesting fluorescent characteristics of QBAs - macarpine (MA), sanguirubine (SR), chelirubine (CHR), sanguilutine (SL), chelilutine (CHL), sanguinarine (SA) and chelerythrine (CHE) upon interacting with living cells. Methods: Water stock solutions of the alkaloids (10-100 mg/ml) were added to intact cells and upon brief incubation the alkaloids stain the cells. Human cell lines HL60 (human promyelocytic leukemia), HeLa (human cervix adenocarcinoma) and LEP (human lung fibroblasts) and piglet blood were used in experiments. Blood cells were stained with MA in a combination with FITC-conjugated anti CD45 surface marker antibody. Cells were analysed by fluorescence microscopy and by flow cytometry. Results: All tested alkaloids immediately enter living cells and MA, CHR and SA bound DNA. MA showed the best DNA staining properties. Fluorescence microscopy of MA, CHR and SA stained cells reveals nuclear architecture and clearly defines chromosomes and apoptotic fragments in living cells. Moreover MA can rapidly report the cellular DNA content of living cells at a resolution level adequate for cell cycle analysis. QBAs were excitable by common argon lasers (488 nm) emitting at the range 575 - 755 nm (i.e. fluorescence detectors FL2-5). Spectral characteristics of MA allow simultaneous surface immunophenotyping. Conclusions: It was proven that MA, CHR and SA stain nucleic acids in living cells. They can be used as supravital fluorescent DNA probes both in fluorescence microscopy and flow cytometry including multiparameter analysis of peripheral blood and bone marrow. MA binds DNA stochiometrically and can report the DNA content.
Abstract (in Czech)
Práce se zabývá možností využití kvartérních benzofenantridinových alkaloidů jako supravitálních fluorescenčních sond zobrazujících DNA.
GA525/04/0017, research and development projectName: Rostlinné zdroje benzofenanthridinových alkaloidů a účinek těchto alkaloidů na normální a nádorové buňky
Investor: Czech Science Foundation, Plant sources of benzophenanthridine alkaloids and effects of these alkaloid on normal and cancer cells
LC06077, research and development projectName: Centrum chemické genetiky
Investor: Ministry of Education, Youth and Sports of the CR
MSM0021622415, plan (intention)Name: Molekulární podstata buněčných a tkáňových regulací
Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations
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