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Vincristine-resistant human laryngeal carcinoma cells demonstrate increased Rous sarcoma virus promoter activity

Dragomira Majhen1, Anamaria Brozovic, Tvrtko Buger

  • 1Laboratory for Genotoxic Agents, Division of Molecular Biology, Ruđer Bošković Institute, 10000 Zagreb, Croatia.

Life Sciences
|September 15, 2010
PubMed
Abstract

Insights

Vincristine-resistant cancer cells show enhanced gene expression from adenovirus vectors. This is due to increased promoter activity, suggesting drug resistance can alter gene therapy effectiveness.

Area of Science:

  • Molecular Biology
  • Gene Therapy
  • Oncology

Background:

  • Gene therapy offers potential for treating drug-resistant cancers.
  • Adenovirus-mediated gene delivery is a key strategy in cancer gene therapy.
  • Understanding transgene expression in resistant cells is crucial for therapeutic development.

Purpose of the Study:

  • To investigate Ad5RSVβgal transgene expression in vincristine-resistant HEp2 cells (VK2).
  • To elucidate the mechanisms underlying altered transgene expression in drug-resistant cancer cells.
  • To assess the role of promoter activity in gene therapy for resistant tumors.

Main Methods:

  • Adenovirus-mediated transgene expression assessed via β-gal staining in HEp2 and VK2 cells.
  • Semiquantitative PCR used to analyze adenovirus attachment and internalization.
  • Reporter gene assays and RT-PCR measured promoter activity following plasmid transfection.

Main Results:

  • VK2 cells demonstrated higher Ad5RSVβgal transgene expression compared to HEp2 cells.
  • Increased expression was observed independently of the coxsackie-adenovirus receptor (CAR).
  • RSV promoter activity was significantly elevated in VK2 cells (33-4.7 fold).

Conclusions:

  • Enhanced transgene expression in VK2 cells is attributed to increased RSV promoter activity.
  • Drug resistance in cancer may correlate with altered promoter activity.
  • Optimizing promoter selection can enhance gene therapy efficacy and reduce vector dosage.

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