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Mid-region parathyroid hormone-related protein (PTHrP) binds chromatin of MDA-MB231 breast cancer cells and isolated oligonucleotides “in vitro”

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Abstract

We have previously shown that PTHrP(38–94)-amide restrains growth and invasion “in vitro”, causes striking toxicity and accelerates death of some breast cancer cell lines, the most responsive being MDA-MB231 whose tumorigenesis was also attenuated “in vivo”. PTHrP(38–94)-amide contains the domain implicated in the nuclear import of PTHrP. Although the nucleus was identified as a destination for mid-region PTHrP, evidence for direct DNA-binding capability is lacking to date. Here, we examined the localization of PTHrP(38–94)-amide within MDA-MB231 cells and within metaphase spread preparations and characterized its DNA-binding properties, employing a combination of immunocytochemical, cytogenetic, “whole genome”/conventional PCR, EMSA and DNase footprinting techniques. The results obtained: (i) show that PTHrP(38–94)-amide gains access to the nuclear compartment of MDA-MB231 cell; (ii) demonstrate that PTHrP(38–94)-amide is a DNA-binding peptide; and, (iii) represent the first data to date on the potential molecular targets in both cellular chromatin and isolated oligonucleotides “in vitro”.

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Abbreviations

PTHrP:

parathyroid hormone-related protein

PTH:

parathyroid hormone

NTS:

nuclear/nucleolar targeting signal

PCR:

polymerase chain reaction

EMSA:

electrophoretic mobility shift assay

BSA:

bovine serum albumin

PBS:

phosphate buffered saline

FITC:

fluorescein isothiocyanate

TBE:

Tris-borate-EDTA

20-m ODN:

20-mer oligonucleotide

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Acknowledgments

This work was supported by a grant from the University of Palermo (R.S. ex 60%) to C.L. and G.B. We thank Prof. A.F. Stewart (University of Pittsburgh, USA) for generous gift of purified PTHrP(38–94)-amide and for helpful suggestions and critical reading of this manuscript

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  1. Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Palermo, Viale delle Scienze, 90128, Palermo, Italy

    Rosalia Sirchia, Marcella Priulla, Giulia Sciandrello, Fabio Caradonna, Giuseppa Barbata & Claudio Luparello

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  1. Rosalia Sirchia
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  2. Marcella Priulla
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  3. Giulia Sciandrello
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  6. Claudio Luparello
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Correspondence to Claudio Luparello.

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Sirchia, R., Priulla, M., Sciandrello, G. et al. Mid-region parathyroid hormone-related protein (PTHrP) binds chromatin of MDA-MB231 breast cancer cells and isolated oligonucleotides “in vitro”. Breast Cancer Res Treat 105, 105–116 (2007). https://doi.org/10.1007/s10549-006-9437-z

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