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Prunetin inhibits nitric oxide activity and induces apoptosis in urinary bladder cancer cells via CASP3 and TNF-α genes

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Abstract

Background

Urinary bladder cancer (UBC) is considered one of the most prevalent malignant tumors worldwide. Complementary and integrative approaches for the treatment of bladder cancer, such as the intake of isoflavonoid phytoestrogens, are of increasing interest due to the risk of mortality and long-term morbidity associated with surgical procedures. The biological effects of prunetin, one of the less-studied phytoestrogens, have not yet been examined in this respect. Therefore, this study aimed to explore the efficacy of prunetin on UBC cells (RT-4).

Methods and results

The cytotoxicity and nitric oxide synthase activities of prunetin were determined in cell cultures. The expression of apoptosis-related genes was determined with RT-PCR. Cell cycle assays were performed using a flow cytometer and cellular apoptotic rate was measured. The results suggested that prunetin has cytotoxic effects at 21.11 µg/mL on RT-4 cells. Flow cytometry analysis showed that prunetin induced apoptosis and arrested th cell cycle in the G0/G1 phase. Prunetin exposure was associated with increases in CASP3 and TNF-α gene expression in RT-4 cells at doses of 21.11 and 42.22 µg/mL, respectively. Strong nitric oxide inhibition was observed at IC50 of 5.18 µg/mL under macrophage mediated inflammatory circumstances.

Conclusions

Prunetin possesses anti-cancer properties and may be a candidate compound for the prevention of UBC. This is the first study that evaluated prunetin for its in vitro antitumor activities, clarified its possible apoptotic molecular mechanism and provided novel insights into its anti-inflammatory nature and effects on the expression of related key genes.

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Acknowledgements

The authors would like to thank Prof. Dr. Anne FRARY from Department of Molecular Biology & Genetics, Izmir Institute of Technology, Izmir, Turkey, for providing kind advice during the drafting of this manuscript. This paper has been submitted to a preprint platform with https://doi.org/10.21203/rs.3.rs-445738/v1.

Funding

This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Grant No. 119Z028).

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Ege University, 35100, Izmir, Turkey

    Çinel Köksal Karayildirim & N. Ülkü Karabay Yavaşoğlu

  2. Department of Bioengineering, Faculty of Engineering, Ege University, 35100, Izmir, Turkey

    Ayşe Nalbantsoy

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  1. Çinel Köksal Karayildirim
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Contributions

ÇKK designed and performed the study and wrote the manuscript. AN prepared the cells for cell culture and performed the cytotoxcity test. NUKY performed the statistical analysis and interpreted data. All authors reviewed the manuscript.

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Correspondence to Çinel Köksal Karayildirim.

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Ethical approval was not required because the experiments utilized in vitro cell culture techniques.

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Köksal Karayildirim, Ç., Nalbantsoy, A. & Karabay Yavaşoğlu, N.Ü. Prunetin inhibits nitric oxide activity and induces apoptosis in urinary bladder cancer cells via CASP3 and TNF-α genes. Mol Biol Rep 48, 7251–7259 (2021). https://doi.org/10.1007/s11033-021-06719-w

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