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PRIMA-1, a mutant p53 reactivator, induces apoptosis and enhances chemotherapeutic cytotoxicity in pancreatic cancer cell lines

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Summary

TP53 mutation is a common event in many cancers, including pancreatic adenocarcinoma, where it occurs in 50–70 % of cases. In an effort to reactivate mutant p53 protein, several new drugs are being developed, including PRIMA-1 and PRIMA-1Met/APR-246 (p53 reactivation and induction of massive apoptosis). PRIMA-1 has been shown to induce apoptosis in tumor cells by reactivating p53 mutants, but its effect in pancreatic cancer remains unclear. Here we investigated the effects of PRIMA-1 on cell viability, cell cycle and expression of p53-regulated proteins in PANC-1 and BxPC-3 (mutant TP53), and CAPAN-2 (wild-type TP53) pancreatic cell lines. Treatment with PRIMA-1 selectively induced apoptosis and cell cycle arrest in p53 mutant cells compared to CAPAN-2 cells. The growth suppressive effect of PRIMA-1 was markedly reduced in p53 mutant cell lines transfected with p53 siRNA, supporting the role of mutant p53 in PRIMA-1 induced cell death. Moreover, treatment with the thiol group donor N-acetylcysteine completely blocked PRIMA-1-induced apoptosis and reinforced the hypothesis that thiol modifications are important for PRIMA-1 biological activity. In combination treatments, PRIMA-1 enhanced the anti-tumor activity of several chemotherapic drugs against pancreatic cancer cells and also exhibited a pronounced synergistic effect in association with the Mdm2 inhibitor Nutlin-3. Taken together, our data indicate that PRIMA-1 induces apoptosis in p53 mutant pancreatic cancer cells by promoting the re-activation of p53 and inducing proapoptotic signaling pathways, providing in vitro evidence for a potential therapeutic approach in pancreatic cancer.

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Acknowledgments

This work was supported by CNPq/Rede Genoprot (Grant #559814/2009-7), CAPES (Brazil) and the International Agency for Research on Cancer (IARC/WHO). Patricia Ashton-Prolla, Guido Lenz, Rafael Roesler and Gilberto Schwartsmann are investigators of CNPq. Patricia Izetti received a grant from CAPES and Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Federal do Rio Grande do Sul, Brazil.

We thank Ms. Ghyslaine Martel-Planche from International Agency for Research on Cancer (IARC) for help with sequencing, Ms. Marie-Piere Cros from IARC for excellent technical assistance during imunofluorescence studies, Rio de Janeiro Cell Bank for assistance with acquisition and establishment of the cell lines in culture and Eduardo Chiela from Universidade Federal do Rio Grande do Sul for help with Western Blotting experiments.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratório de Medicina Genômica, Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Patricia Izetti, Juliana Giacomazzi, Bárbara Alemar & Patricia Ashton-Prolla

  2. Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500–Prédio 43323M, Porto Alegre, RS, Brazil, 91501-970

    Patricia Izetti, Bárbara Alemar & Patricia Ashton-Prolla

  3. Laboratório de Pesquisas em Câncer, Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Patricia Izetti, Ana Lucia Abujamra, Caroline Brunetto de Farias, Rafael Roesler & Gilberto Schwartsmann

  4. International Agency for Research on Cancer, 150 Cours Albert Thomas, Lyon, France, 69372

    Agnes Hautefeuille

  5. Instituto do Câncer Infantil do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Ana Lucia Abujamra & Caroline Brunetto de Farias

  6. Instituto Nacional de Ciência e Tecnologia Translacional em Medicina, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Ana Lucia Abujamra, Caroline Brunetto de Farias, Rafael Roesler & Gilberto Schwartsmann

  7. Laboratório de Sinalização e Plasticidade Celular, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500–Prédio 43431, Porto Alegre, RS, Brazil, 91501-970

    Guido Lenz

  8. Laboratório de Neurofarmacologia e Biologia de Tumores Neurais, Departamento de Farmacologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500–Sala 211, Porto Alegre, RS, Brazil, 91501-970

    Rafael Roesler

  9. Serviço de Oncologia Clínica, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Gilberto Schwartsmann

  10. Serviço de Cirurgia do Aparelho Digestivo, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil, 90035-003

    Alessandro Bersch Osvaldt

  11. International Prevention Research Institute, 95 Cours Lafayette, Lyon, France, 69006

    Pierre Hainaut

  12. Departamento de Genética, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500–Prédio 43323M, Porto Alegre, RS, Brazil, 91501-970

    Patricia Ashton-Prolla

  13. Laboratório de Medicina Genômica, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, 90035-903, Porto Alegre, RS, Brazil

    Patricia Izetti

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  1. Patricia Izetti
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  2. Agnes Hautefeuille
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  3. Ana Lucia Abujamra
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  4. Caroline Brunetto de Farias
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  10. Alessandro Bersch Osvaldt
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  11. Pierre Hainaut
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  12. Patricia Ashton-Prolla
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Correspondence to Patricia Izetti.

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Izetti, P., Hautefeuille, A., Abujamra, A.L. et al. PRIMA-1, a mutant p53 reactivator, induces apoptosis and enhances chemotherapeutic cytotoxicity in pancreatic cancer cell lines. Invest New Drugs 32, 783–794 (2014). https://doi.org/10.1007/s10637-014-0090-9

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  • DOI: https://doi.org/10.1007/s10637-014-0090-9

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