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Gemcitabine/Cisplatin Treatment Induces Concomitant SERTAD1, CDKN2B and GADD45A Modulation and Cellular Changes in Bladder Cancer Cells Regardless of the Site of TP53 Mutation

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Pathology & Oncology Research

Abstract

Simultaneous use of cisplatin (CIS) and gemcitabine (GEN) for treating bladder cancer has increased because of their complementary effects. However, the molecular mechanisms underlying the activities of these two antineoplastic drugs are not fully known. Here, molecular biology techniques and microscopy were used to investigate transcriptomic and morphological changes in low and high-grade urinary bladder transitional carcinoma cell lines [RT4 - wild type TP53; 5637 - two TP53 mutations, one in codon 72 (Arg-Pro) and other in codon 280 (Arg-Thr) and T24 - in-frame deletion of tyrosine 126 in the TP53 allele] simultaneously treated with CIS/GEN. Gene expression profile was evaluated by PCR arrays; cell morphology by scanning and transmission electron microscopy, and apoptosis was analyzed using fluorescent dye. Results showed concomitantly upregulation of CDKN2B (G1/S transition), GADD45A (DNA repair and apoptosis) and SERTAD1 (regulation of transcription) gene, increased number of nuclear chamfers and apoptotic cells, and reduced number of microfilaments, organelles and in the size of the nucleus in 5637 and T24 cells after simultaneous treatment with CIS/GEN. In conclusion, independently of the TP53 mutation status and tumor grade, CIS/GEN induced gene modulation accompanied by changes in cell morphologies, which confirm the antiproliferative activity of the treatment protocol. These findings help to understand the pathways modulated by these antineoplastic agents and may provide insights for anti-cancer chemotherapy.

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Authors and Affiliations

  1. School of Pharmacy, UFOP – Federal University of Ouro Preto, Ouro Preto, MG, Brazil

    Glenda Nicioli da Silva

  2. Escola de Farmácia, Departamento de Análises Clínicas, UFOP – Universidade Federal de Ouro Preto, Ouro Preto, MG, 35400-000, Brazil

    Glenda Nicioli da Silva

  3. Institute of Physics, USP – University of São Paulo, São Paulo, SP, Brazil

    Leandro Toshio Filoni & Maria Cecília Salvadori

  4. Medical School, UNESP – State University of São Paulo, Botucatu, SP, Brazil

    Daisy Maria Fávero Salvadori

Authors
  1. Glenda Nicioli da Silva
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  2. Leandro Toshio Filoni
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  3. Maria Cecília Salvadori
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  4. Daisy Maria Fávero Salvadori
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Contributions

GNS contributed to the experimental design and the interpretation of data, performed the scanning and transmission electron microscopy and PCR array experiments, and wrote the manuscript. LTF performed scanning electron microscopy. MCBS contributed to the interpretation of the data and critically read the manuscript. DMFS contributed to the experimental design, the interpretation of data, and the critical reading of the manuscript.

Corresponding author

Correspondence to Glenda Nicioli da Silva.

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The authors declare that they have no competing interests.

Funding

This study was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP - 2008/09147–6), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (CBB - APQ-01497-14) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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da Silva, G.N., Filoni, L.T., Salvadori, M.C. et al. Gemcitabine/Cisplatin Treatment Induces Concomitant SERTAD1, CDKN2B and GADD45A Modulation and Cellular Changes in Bladder Cancer Cells Regardless of the Site of TP53 Mutation. Pathol. Oncol. Res. 24, 407–417 (2018). https://doi.org/10.1007/s12253-017-0255-x

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  • DOI: https://doi.org/10.1007/s12253-017-0255-x

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