Inhibition of DNA polymerase α expression and cell growth, a possible triple helix mechanism

Angela Alama, Federica Barbieri, Monica Cagnoli, Mauro Mazzei, Teresa Grandi, Angelo Nicolin

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The antiproliferative effects mediated by a 14-mer homopyrimidine oligonucleotide (5' CTTTCTCTTTTCTC3' designed to form DNA triplex with a purine region of the DNA polymerase α promoter, were evaluated on the human breast cancer cell line MDA-MB 231. In order to stabilize the triplex complex under physiologic conditions, replacement of cytosines by methylcytosines in the oligomer sequence was carried out. Band-shift analyses demonstrated a complete triplex formation between the radiolabeled target duplex DNA and the methylcytosine-modified oligomer at the concentration of 0.1 μM under physiologic pH and temperature. A single exposure of MDA-MB 231 cells to 0.5 μM methylcytosine-modified oligonucleotide was able to markedly reduce the cell number and the percentage of cells in DNA synthesis up to 58% and 66%, respectively, compared with controls. Furthermore, a 48% reduction in the amount of the DNA polymerase a mRNA was reported after treatment with the oligomer. In conclusion, data from the present study demonstrate that an oligonucleotide to DNA polymerase α promoter, designed to form a triple helix with target double-stranded DNA, inhibits the expression of the reporter gene at the biologic and molecular levels, suggesting a possible triplex-mediated mechanism of action.

Original languageEnglish
Pages (from-to)95-101
Number of pages7
JournalAntisense and Nucleic Acid Drug Development
Volume6
Issue number2
Publication statusPublished - 1996

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DNA-Directed DNA Polymerase
Oligonucleotides
DNA
Growth
Cytosine
Reporter Genes
Cell Count
Breast Neoplasms
Cell Line
Messenger RNA
Temperature

ASJC Scopus subject areas

  • Genetics
  • Pharmacology

Cite this

Inhibition of DNA polymerase α expression and cell growth, a possible triple helix mechanism. / Alama, Angela; Barbieri, Federica; Cagnoli, Monica; Mazzei, Mauro; Grandi, Teresa; Nicolin, Angelo.

In: Antisense and Nucleic Acid Drug Development, Vol. 6, No. 2, 1996, p. 95-101.

Research output: Contribution to journalArticle

Alama, A, Barbieri, F, Cagnoli, M, Mazzei, M, Grandi, T & Nicolin, A 1996, 'Inhibition of DNA polymerase α expression and cell growth, a possible triple helix mechanism', Antisense and Nucleic Acid Drug Development, vol. 6, no. 2, pp. 95-101.
Alama, Angela ; Barbieri, Federica ; Cagnoli, Monica ; Mazzei, Mauro ; Grandi, Teresa ; Nicolin, Angelo. / Inhibition of DNA polymerase α expression and cell growth, a possible triple helix mechanism. In: Antisense and Nucleic Acid Drug Development. 1996 ; Vol. 6, No. 2. pp. 95-101.
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