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Effects in live cells of a c-myc anti-gene PNA linked to a nuclear localization signal

Nature Biotechnology volume 18, pages 300–303 (2000)Cite this article

Abstract

Peptide nucleic acids (PNA) are synthetic homologs of nucleic acids in which the phosphate-sugar polynucleotide backbone is replaced by a flexible polyamide. In this study, a PNA construct was employed as an anti-gene agent in intact cells in culture. The cell lines studied were derived from Burkitt's lymphomas (BL) that presented a translocated and hyperexpressed c-myc oncogene. A 17-mer anti-myc PNA, complementary to a unique sequence located at the beginning of the second exon of the oncogene, and was covalently linked at its N terminus to a nuclear localization signal (NLS) (PNA-mycwt-NLS). When BL cells were exposed to PNA-mycwt-NLS, the anti-gene construct was localized predominantly in the cell nuclei and a rapid consequent downregulation of c-myc expression occurred. Under these conditions, both completion of a productive cell cycle and apoptosis were inhibited.

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Figure 1: Delivery of PNA-NLS to intact nuclei of BL cells.
Figure 2: Inhibition of c-myc expression by PNA-mycwt-NLS.
Figure 3: Growth inhibition of HBL2 cell line by PNA-mycwt-NLS.
Figure 4: Cell death induced by PNAs is unrelated to apoptosis.
Figure 5: Apoptosis prevention in BL cells exposed to F(ab′)2 α-μ-ab.

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Acknowledgements

We thank Prof. A. Alagòn (Instituto de Biotecnologia, UNAM, Cuernavaca, MX) and Prof. E. Melloni (Department of Experimental Medicine, Biochemistry Section, University of Genoa, Italy) for permission to use their microscopy facilities; X. Alvarado, A. Olvera, and Dr. M. Passalacqua for performing the confocal microscopy; and Dr. R. Stock for assistance in image analysis. This work was supported by: Associazione Italiana per la Ricerca sul Cancro (AIRC) and by CNR/CONACYT to L.C.B.; Istituto Superiore di Sanità (ISS) AIDS Project and AIRC to M.F.

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Authors and Affiliations

  1. Servizi di Immunologia Clinica, National Cancer Institute, Genoa, Italy

    Giovanna Cutrona, Massimo Ulivi, Silvio Roncella & Manlio Ferrarini

  2. Servizi di Oncologia Sperimentale, National Cancer Institute, Genoa, Italy

    Elisabetta M. Carpaneto & Lidia C. Boffa

  3. Servizio di Anatomia Patologica, Ospedale Sant'Andrea, La Spezia, Italy

    Silvio Roncella

  4. TIB MOLBIOL, Berlin, D-10829, Germany

    Olfert Landt

  5. Dipartimento di Oncologia, Biologia e Genetica, Università degli Studi di Genova, Genoa, Italy

    Manlio Ferrarini

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  1. Giovanna Cutrona
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Correspondence to Lidia C. Boffa.

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Cutrona, G., Carpaneto, E., Ulivi, M. et al. Effects in live cells of a c-myc anti-gene PNA linked to a nuclear localization signal. Nat Biotechnol 18, 300–303 (2000). https://doi.org/10.1038/73745

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