Fisetin induces DNA double-strand break and interferes with the repair of radiation-induced damage to radiosensitize triple negative breast cancer cells.

Autor: Khozooei S; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany., Lettau K; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany., Barletta F; Quantitative Biology Center (QBiC), University of Tuebingen, Auf der Morgenstelle 10, 72076, Tuebingen, Germany., Jost T; Department of Radiation Oncology, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.; Comprehensive Cancer Center Erlangen-EMN (CCC ER-EMN), Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany., Rebholz S; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany., Veerappan S; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany., Franz-Wachtel M; Proteome Center Tuebingen, University of Tuebingen, Tuebingen, Germany., Macek B; Proteome Center Tuebingen, University of Tuebingen, Tuebingen, Germany., Iliakis G; Divison of Experimental Radiation Oncology, Department of Radiation Oncology, Institutsgruppe 1, Bauteil A, 4.038, University of Duisburg-Essen, Hufelandstr. 55, 45122 , Essen, Germany., Distel LV; Department of Radiation Oncology, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.; Comprehensive Cancer Center Erlangen-EMN (CCC ER-EMN), Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany., Zips D; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany., Toulany M; Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, University of Tuebingen, Roentgenweg 11, 72076, Tuebingen, Germany. mahmoud.toulany@uni-tuebingen.de.; German Cancer Consortium (DKTK), partner site Tuebingen, German Cancer Research Center (DKFZ), Heidelberg, Germany. mahmoud.toulany@uni-tuebingen.de.
Jazyk: angličtina
Zdroj: Journal of experimental & clinical cancer research : CR [J Exp Clin Cancer Res] 2022 Aug 22; Vol. 41 (1), pp. 256. Date of Electronic Publication: 2022 Aug 22.
DOI: 10.1186/s13046-022-02442-x
Abstrakt: Background: Triple-negative breast cancer (TNBC) is associated with aggressiveness and a poor prognosis. Besides surgery, radiotherapy serves as the major treatment modality for TNBC. However, response to radiotherapy is limited in many patients, most likely because of DNA damage response (DDR) signaling mediated radioresistance. Y-box binding protein-1 (YB-1) is a multifunctional protein that regulates the cancer hallmarks among them resisting to radiotherapy-induced cell death. Fisetin, is a plant flavonol of the flavonoid family of plant polyphenols that has anticancer properties, partially through inhibition of p90 ribosomal S6 kinase (RSK)-mediated YB-1 phosphorylation. The combination of fisetin with radiotherapy has not yet been investigated.
Methods: Activation status of the RSK signaling pathway in total cell lysate and in the subcellular fractions was analyzed by Western blotting. Standard clonogenic assay was applied to test post-irradiation cell survival. γH2AX foci assay and 3 color fluorescence in situ hybridization analyses were performed to study frequency of double-strand breaks (DSB) and chromosomal aberrations, respectively. The underlying repair pathways targeted by fisetin were studied in cells expressing genomically integrated reporter constructs for the DSB repair pathways via quantifying the expression of green fluorescence protein by flow cytometry. Flow cytometric quantification of sub-G1 cells and the protein expression of LC3-II were employed to measure apoptosis and autophagy, respectively. Kinase array and phosphoproteomics were performed to study the effect of fisetin on DDR response signaling.
Results: We showed that the effect of fisetin on YB-1 phosphorylation in TNBC cells is comparable to the effect of the RSK pharmacological inhibitors. Similar to ionizing radiation (IR), fisetin induces DSB. Additionally, fisetin impairs repair of IR-induced DSB through suppressing the classical non-homologous end-joining and homologous recombination repair pathways, leading to chromosomal aberration as tested by metaphase analysis. Effect of fisetin on DSB repair was partially dependent on YB-1 expression. Phosphoproteomic analysis revealed that fisetin inhibits DDR signaling, which leads to radiosensitization in TNBC cells, as shown in combination with single dose or fractionated doses irradiation.
Conclusion: Fisetin acts as a DSB-inducing agent and simultaneously inhibits repair of IR-induced DSB. Thus, fisetin may serve as an effective therapeutic strategy to improve TNBC radiotherapy outcome.
(© 2022. The Author(s).)
Databáze: MEDLINE
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