Quantitative approaches for characterizing molecular organization of cell membrane molecules under physiological and pathological conditions profit from recently developed super-resolution imaging techniques. Current tools employ statistical algorithms to determine clusters of molecules based on single-molecule localization microscopy (SMLM) data. These approaches are limited by the ability of SMLM techniques to identify and localize molecules in densely populated areas and experimental conditions of sample preparation and image acquisition. We have developed a robust, model-free, quantitative clustering analysis to determine the distribution of membrane molecules that excels in densely labeled areas and is tolerant to various experimental conditions, i.e. multiple-blinking or high blinking rates. The method is based on a TIRF microscope followed by a super-resolution optical fluctuation imaging (SOFI) analysis. The effectiveness and robustness of the method is validated using simulated and experimental data investigating nanoscale distribution of CD4 glycoprotein mutants in the plasma membrane of T cells.

• MeSH
• algoritmy MeSH
• antigeny CD4 genetika   metabolismus   MeSH
• buněčná membrána metabolismus   MeSH
• fluorescenční barviva MeSH
• fluorescenční mikroskopie metody   statistika a číselné údaje   MeSH
• Jurkat buňky MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• mutantní proteiny genetika   metabolismus   MeSH
• optické zobrazování metody   statistika a číselné údaje   MeSH
• shluková analýza MeSH
• T-lymfocyty imunologie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• validační studie MeSH

Nordamnacanthal, an anthraquinone extracted from the root of Morinda elliptica from Rubiaceae family has cytotoxic properties towards cancer cell lines and antitumor promoting activities. This study was conducted to determine the cytotoxic effect of nordamnacanthal towards MOLT-4 and MCF-7 cell lines. Nordamnacanthal was found to be more cytotoxic towards MOLT-4 than MCF-7 with the IC50 of 3.8 μg/ml and 54 μg/ml, respectively, as detected by using the trypan blue dye exclusion test. The nordamnacanthal-treated cells showed characteristics of apoptosis such as membrane blebbing, chromatin condensation and formation of apoptotic bodies as observed under an inverted light microscope. Fluorescence analysis of cell death using acridine orange and propidium iodide staining showed that the population of MOLT-4 and MCF-7 cells underwent apoptosis at the IC50 value was 32% and 30.4%, respectively. Cell cycle analysis by flow cytometry indicated that nordamnacanthal did arrest MCF-7 cells at the G2/M phase. For MOLT-4, no cell cycle arrest was observed. Bcl-2 and Bax were downregulated in nordamnacanthal-treated MCF-7 cells. On the other hand, expression of the proteins in MOLT-4 was not significantly different from the control. In conclusion, nordamnacanthal was more cytotoxic towards MOLT-4 than MCF-7 cell line. The compound induced apoptosis in both cell lines, but with G2/M arrest and the involvement of Bcl-2 and Bax only in MCF-7.

• Klíčová slova
• cytotoxicita,
• MeSH
• adenokarcinom patologie   MeSH
• aldehydy chemie   terapeutické užití   MeSH
• anthrachinony * chemie   terapeutické užití   MeSH
• apoptóza * účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• cytotoxiny MeSH
• down regulace MeSH
• ELISA statistika a číselné údaje   MeSH
• fluorescenční barviva MeSH
• fluorescenční mikroskopie statistika a číselné údaje   MeSH
• inhibiční koncentrace 50 MeSH
• kontrolní body fáze G2 buněčného cyklu účinky léků   MeSH
• kořeny rostlin MeSH
• kultivované buňky MeSH
• lidé MeSH
• lymfoblastická leukemie-lymfom z prekurzorových T-buněk patologie   MeSH
• MFC-7 buňky cytologie   účinky léků   MeSH
• Morinda MeSH
• nádorové buněčné linie * cytologie   účinky léků   MeSH
• nádory prsu * patologie   MeSH
• protein X asociovaný s bcl-2 MeSH
• průtoková cytometrie statistika a číselné údaje   MeSH
• rostlinné extrakty MeSH
• statistika jako téma MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH

• MeSH
• buněčné jádro genetika   účinky záření   ultrastruktura   MeSH
• buněčný cyklus fyziologie   MeSH
• chromatin fyziologie   účinky záření   ultrastruktura   MeSH
• fluorescenční mikroskopie metody   statistika a číselné údaje   MeSH
• hybridizace in situ fluorescenční metody   statistika a číselné údaje   MeSH
• lidé MeSH
• nádorové buňky kultivované fyziologie   ultrastruktura   MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH