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Cancer gene therapy mediated by RALA/plasmid DNA vectors: Nitrogen to phosphate groups ratio (N/P) as a tool for

A R Neves1, A Sousa1, R Faria1

  • 1CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.

Colloids and Surfaces. B, Biointerfaces
|November 13, 2019
PubMed

Insights

This study introduces RALA peptide as a carrier for p53 plasmid DNA (pDNA) delivery, enhancing cancer gene therapy. The nitrogen to phosphate (N/P) ratio effectively controls p53 protein expression and cancer cell apoptosis.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cancer Research

Background:

  • Cancer gene therapy using the p53 tumor suppressor gene is promising but requires effective delivery systems.
  • Cell-penetrating peptides offer potential for improved therapeutic efficacy in gene delivery.

Purpose of the Study:

  • To explore the RALA peptide's ability to condense p53 plasmid DNA (pDNA) for intracellular delivery.
  • To investigate the RALA/pDNA complex as a platform for p53 gene therapy in cancer.

Main Methods:

  • Characterization of RALA/pDNA complexes (nanoparticles) including morphology, size, charge, and loading capacity.
  • Fourier-transformed infrared (FTIR) spectroscopy for structural analysis.
  • Confocal microscopy for intracellular localization and uptake studies.
  • In vitro transfection of HeLa cells to assess gene release, p53 protein expression, and apoptosis induction.

Main Results:

  • RALA peptide effectively condensed pDNA, forming nanoparticles suitable for intracellular delivery.
  • Confocal microscopy confirmed nanoparticle uptake and sustained pDNA delivery.
  • In vitro studies demonstrated RALA/pDNA mediated p53 gene expression and subsequent apoptosis in HeLa cells.
  • The nitrogen to phosphate (N/P) ratio was identified as a critical factor modulating transfection efficiency, protein expression, and apoptosis.

Conclusions:

  • RALA peptide-based delivery systems are effective for p53 gene therapy, enabling sustained pDNA uptake and expression.
  • The N/P ratio serves as a tunable parameter to optimize RALA/pDNA vector properties and therapeutic outcomes.
  • This approach holds significant potential for advancing p53-mediated cancer therapy by controlling gene delivery and apoptosis induction.
Keywords:
ApoptosisCancer gene therapyIn vitro transfectionN/P ratioRALA peptideRALA/pDNA vectorsp53 genep53 protein

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