TY  - JOUR
AU  - Kucirkova, Tereza
AU  - Stiborek, Marek
AU  - Ducka, Monika
AU  - Navratilova, Jarmila
AU  - Bogdanović Pristov, Jelena
AU  - Popovic-Bijelic, Ana
AU  - Vojvodić, Snežana
AU  - Preisler, Jan
AU  - Kanicky, Viktor
AU  - Smarda, Jan
AU  - Spasojević, Ivan
AU  - Benes, Petr
PY  - 2018
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/1156
AB  - Wedelactone (WL), a plant polyphenolic derivative of coumestan, represents a promising anti-cancer agent. The underlying mechanisms of its action are not fully understood and appear to involve interplay with copper ions. Herein, we examined coordination and redox interactions of WL with Cu2+ in phosphate buffer (pH 7), and in two breast cancer cell lines. EPR, UV-Vis and fluorescence spectroscopy showed that WL and Cu2+ build a coordination complex with 2:1 stoichiometry and distorted tetrahedral geometry. WL showed strong fluorescence that was quenched by Cu2+. The sequestration of the intracellular copper pool with neocuproine led to a significant drop in the cytotoxic effects of WL, whereas the co-application of Cu2+ and WL and the formation of an extracellular complex suppressed both the cytotoxic effects of WL and copper loading. Fluorescence microscopy showed that WL is mainly localized in the cytosol and significantly less in the nuclei. WL fluorescence was stronger in cells pretreated with neocuproine, implying that the complex of WL and Cu2+ is formed inside the cells. WL caused a two-fold increase in the lysosomal level of copper as well as copper-dependent lysosome membrane permeabilization. On the other hand, the protective effects of overexpression of thioredoxin 1 imply that WL exerts the main oxidative impact inside the nucleus. The interactions of WL with copper may be essential for therapeutic performance and selectivity against cancer cells, taking into account that a number of cancer types, including breast cancer, exhibit increased intratumoral copper levels or altered copper distribution.
PB  - Royal Soc Chemistry, Cambridge
T2  - Metallomics
T1  - Anti-cancer effects of wedelolactone: interactions with copper and subcellular localization
EP  - 1531
IS  - 10
SP  - 1524
VL  - 10
DO  - 10.1039/c8mt00191j
ER  - 

@article{
author = "Kucirkova, Tereza and Stiborek, Marek and Ducka, Monika and Navratilova, Jarmila and Bogdanović Pristov, Jelena and Popovic-Bijelic, Ana and Vojvodić, Snežana and Preisler, Jan and Kanicky, Viktor and Smarda, Jan and Spasojević, Ivan and Benes, Petr",
year = "2018",
abstract = "Wedelactone (WL), a plant polyphenolic derivative of coumestan, represents a promising anti-cancer agent. The underlying mechanisms of its action are not fully understood and appear to involve interplay with copper ions. Herein, we examined coordination and redox interactions of WL with Cu2+ in phosphate buffer (pH 7), and in two breast cancer cell lines. EPR, UV-Vis and fluorescence spectroscopy showed that WL and Cu2+ build a coordination complex with 2:1 stoichiometry and distorted tetrahedral geometry. WL showed strong fluorescence that was quenched by Cu2+. The sequestration of the intracellular copper pool with neocuproine led to a significant drop in the cytotoxic effects of WL, whereas the co-application of Cu2+ and WL and the formation of an extracellular complex suppressed both the cytotoxic effects of WL and copper loading. Fluorescence microscopy showed that WL is mainly localized in the cytosol and significantly less in the nuclei. WL fluorescence was stronger in cells pretreated with neocuproine, implying that the complex of WL and Cu2+ is formed inside the cells. WL caused a two-fold increase in the lysosomal level of copper as well as copper-dependent lysosome membrane permeabilization. On the other hand, the protective effects of overexpression of thioredoxin 1 imply that WL exerts the main oxidative impact inside the nucleus. The interactions of WL with copper may be essential for therapeutic performance and selectivity against cancer cells, taking into account that a number of cancer types, including breast cancer, exhibit increased intratumoral copper levels or altered copper distribution.",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "Metallomics",
title = "Anti-cancer effects of wedelolactone: interactions with copper and subcellular localization",
pages = "1531-1524",
number = "10",
volume = "10",
doi = "10.1039/c8mt00191j"
}

Kucirkova, T., Stiborek, M., Ducka, M., Navratilova, J., Bogdanović Pristov, J., Popovic-Bijelic, A., Vojvodić, S., Preisler, J., Kanicky, V., Smarda, J., Spasojević, I.,& Benes, P.. (2018). Anti-cancer effects of wedelolactone: interactions with copper and subcellular localization. in Metallomics
Royal Soc Chemistry, Cambridge., 10(10), 1524-1531.
https://doi.org/10.1039/c8mt00191j

Kucirkova T, Stiborek M, Ducka M, Navratilova J, Bogdanović Pristov J, Popovic-Bijelic A, Vojvodić S, Preisler J, Kanicky V, Smarda J, Spasojević I, Benes P. Anti-cancer effects of wedelolactone: interactions with copper and subcellular localization. in Metallomics. 2018;10(10):1524-1531.
doi:10.1039/c8mt00191j .

Kucirkova, Tereza, Stiborek, Marek, Ducka, Monika, Navratilova, Jarmila, Bogdanović Pristov, Jelena, Popovic-Bijelic, Ana, Vojvodić, Snežana, Preisler, Jan, Kanicky, Viktor, Smarda, Jan, Spasojević, Ivan, Benes, Petr, "Anti-cancer effects of wedelolactone: interactions with copper and subcellular localization" in Metallomics, 10, no. 10 (2018):1524-1531,
https://doi.org/10.1039/c8mt00191j . .

TY  - JOUR
AU  - Nehybova, Tereza
AU  - Smarda, Jan
AU  - Daniel, Lukas
AU  - Stiborek, Marek
AU  - Kanicky, Viktor
AU  - Spasojević, Ivan
AU  - Preisler, Jan
AU  - Damborsky, Jiri
AU  - Benes, Petr
PY  - 2017
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/1048
AB  - Wedelolactone is a multi-target natural plant coumestan exhibiting cytotoxicity towards cancer cells. Although several molecular targets of wedelolactone have been recognized, the molecular mechanism of its cytotoxicity has not yet been elucidated. In this study, we show that wedelolactone acts as an inhibitor of chymotrypsin-like, trypsin-like, and caspase-like activities of proteasome in breast cancer cells. The proteasome inhibitory effect of wedelolactone was documented by (i) reduced cleavage of fluorogenic proteasome substrates; (ii) accumulation of polyubiquitinated proteins and proteins with rapid turnover in tumor cells; and (iii) molecular docking of wedelolactone into the active sites of proteasome catalytic subunits. Inhibition of proteasome by wedelolactone was independent on its ability to induce reactive oxygen species production by redox cycling with copper ions, suggesting that wedelolactone acts as copper-independent proteasome inhibitor. We conclude that the cytotoxicity of wedelolactone to breast cancer cells is partially mediated by targeting proteasomal protein degradation pathway. Understanding the structural basis for inhibitory mode of wedelolactone might help to open up new avenues for design of novel compounds efficiently inhibiting cancer cells.
PB  - MDPI, Basel
T2  - International Journal of Molecular Sciences
T1  - Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells
IS  - 4
VL  - 18
DO  - 10.3390/ijms18040729
ER  - 

@article{
author = "Nehybova, Tereza and Smarda, Jan and Daniel, Lukas and Stiborek, Marek and Kanicky, Viktor and Spasojević, Ivan and Preisler, Jan and Damborsky, Jiri and Benes, Petr",
year = "2017",
abstract = "Wedelolactone is a multi-target natural plant coumestan exhibiting cytotoxicity towards cancer cells. Although several molecular targets of wedelolactone have been recognized, the molecular mechanism of its cytotoxicity has not yet been elucidated. In this study, we show that wedelolactone acts as an inhibitor of chymotrypsin-like, trypsin-like, and caspase-like activities of proteasome in breast cancer cells. The proteasome inhibitory effect of wedelolactone was documented by (i) reduced cleavage of fluorogenic proteasome substrates; (ii) accumulation of polyubiquitinated proteins and proteins with rapid turnover in tumor cells; and (iii) molecular docking of wedelolactone into the active sites of proteasome catalytic subunits. Inhibition of proteasome by wedelolactone was independent on its ability to induce reactive oxygen species production by redox cycling with copper ions, suggesting that wedelolactone acts as copper-independent proteasome inhibitor. We conclude that the cytotoxicity of wedelolactone to breast cancer cells is partially mediated by targeting proteasomal protein degradation pathway. Understanding the structural basis for inhibitory mode of wedelolactone might help to open up new avenues for design of novel compounds efficiently inhibiting cancer cells.",
publisher = "MDPI, Basel",
journal = "International Journal of Molecular Sciences",
title = "Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells",
number = "4",
volume = "18",
doi = "10.3390/ijms18040729"
}

Nehybova, T., Smarda, J., Daniel, L., Stiborek, M., Kanicky, V., Spasojević, I., Preisler, J., Damborsky, J.,& Benes, P.. (2017). Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells. in International Journal of Molecular Sciences
MDPI, Basel., 18(4).
https://doi.org/10.3390/ijms18040729

Nehybova T, Smarda J, Daniel L, Stiborek M, Kanicky V, Spasojević I, Preisler J, Damborsky J, Benes P. Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells. in International Journal of Molecular Sciences. 2017;18(4).
doi:10.3390/ijms18040729 .

Nehybova, Tereza, Smarda, Jan, Daniel, Lukas, Stiborek, Marek, Kanicky, Viktor, Spasojević, Ivan, Preisler, Jan, Damborsky, Jiri, Benes, Petr, "Wedelolactone Acts as Proteasome Inhibitor in Breast Cancer Cells" in International Journal of Molecular Sciences, 18, no. 4 (2017),
https://doi.org/10.3390/ijms18040729 . .