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Updated: Sep 29, 2026
Evaluating the Effectiveness of Cancer Drug Sensitization In Vitro and In Vivo
Published on: February 6, 2015
Recombinant adenovirus-mediated p14(ARF) overexpression sensitizes human breast cancer cells to cisplatin
Xiyun Deng1, Min Kim, Didier Vandier
1Medicine Branch, National Cancer Institute, National Institutes of Health, Building 10, Room 10N226, Bethesda, MD 20892, USA.
Abstract:
p14(ARF), the alternative product from the human INK4a/ARF locus, is one of the major targets for alterations in the development of human cancers. Overexpression of p14(ARF) results in cell cycle arrest and apoptosis. To examine the potential therapeutic role of re-expressing p14(ARF) gene product in human breast cancer, a recombinant adenovirus expressing the human p14(ARF) cDNA (Adp14(ARF)) was constructed and used to infect breast cancer cells. Five days after infection, Adp14(ARF) had considerable cytotoxicity on p53-wild-type MCF-7 cells. A time-course study showed that Adp14(ARF) infection of MCF-7 cells at 100pfu/cell increased the number of cells in G0/G1 phase and decreased that in S and G2/M phases. The presence of apoptotic cells was confirmed using the TUNEL assay. Adp14(ARF)-mediated expression of p14(ARF) also resulted in a considerable increase in the amounts of p53 and its target proteins, p21(WAF1) and MDM2. Furthermore, the combination treatment of MCF-7 cells with Adp14(ARF) and cisplatin resulted in a significantly greater cell death. Together, we conclude that p14(ARF) plays an important role in the induction of cell cycle arrest and apoptosis in breast cancer cells and recombinant adenovirus-mediated p14(ARF) expression greatly increases the sensitivity of these cells to cisplatin. These results demonstrate that the proper combination of Adp14(ARF) with conventional chemotherapeutic drug(s) could have potential benefits in treating breast cancer that carries wild-type p53 gene.
Insights
Re-expressing the p14ARF gene using Adp14ARF in breast cancer cells induces cell cycle arrest and apoptosis. This approach enhances sensitivity to cisplatin, suggesting a potential therapeutic strategy for p53-wild-type breast cancers.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- The p14ARF protein, a product of the INK4a/ARF locus, is frequently altered in human cancers.
- Overexpression of p14ARF induces cell cycle arrest and apoptosis, making it a potential therapeutic target.
Purpose of the Study:
- To investigate the therapeutic potential of re-expressing p14ARF in human breast cancer cells.
- To evaluate the efficacy of a recombinant adenovirus carrying the p14ARF gene (Adp14ARF) in breast cancer treatment.
Main Methods:
- Construction of a recombinant adenovirus expressing human p14ARF cDNA (Adp14ARF).
- Infection of p53-wild-type MCF-7 breast cancer cells with Adp14ARF.
- Assessment of cell viability, cell cycle distribution (G0/G1, S, G2/M phases), apoptosis (TUNEL assay), and protein levels (p53, p21WAF1, MDM2).
- Combination treatment with Adp14ARF and cisplatin.
Main Results:
- Adp14ARF demonstrated significant cytotoxicity in MCF-7 cells.
- Adp14ARF infection led to G0/G1 cell cycle arrest and increased apoptosis.
- Expression of p14ARF increased levels of p53, p21WAF1, and MDM2.
- Combined Adp14ARF and cisplatin treatment resulted in significantly greater cell death.
Conclusions:
- p14ARF is crucial for inducing cell cycle arrest and apoptosis in breast cancer cells.
- Adenovirus-mediated p14ARF expression enhances breast cancer cell sensitivity to cisplatin.
- Combination therapy with Adp14ARF and chemotherapy holds promise for treating p53-wild-type breast cancer.
