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Evaluation of a Synthetic Polyethyleneimine Based Polymeric Vector for ING4 Gene Delivery to MCF-7 Breast Cancer
Uğur Karagöz1, Remant Bahadur Kc2, Elif Isel3
1Trakya University Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, Edirne, Türkiye.
Objectives:
Breast cancer is the most common type of cancer among women and the second most common cause of death after lung cancer. The inhibitor of growth (ING) transcript levels are often suppressed in cancer cells, making it a promising candidate for cancer therapy. In this study, we aimed to formulate a polyplex that effectively carries and delivers pING4 to breast cancer cells.
Materials And Methods:
Polyethyleneimine (PEI)-based non-viral vectors were synthesized and characterized for plasmid DNA delivery. Complexation was achieved via electrostatic interactions between the synthesized polymeric vectors and plasmid DNA. Characterization studies were conducted by testing Sodium dodecyl sulfate-induced complexation, Deoxyribonuclease I protection, and serum stability of the polyplexes. Subsequently, polyplexes were tested on MCF-7 cells for anticancer activity using the XTT cell viability assay. Western blot analysis was performed for the ING4 protein.
Results:
Polyplexes carrying the ING4 gene exhibited significantly lower cell viability than control polyplexes (p=0.0067). During the 5-day viability assay, the lowest cell viability was observed on day 4. Approximately 69.11±2.18% cell viability was observed with ING4 treatment and the control group showed no cell death on day 4 (101.53±5.06%). The prepared delivery systems did not show a toxic effect on MCF-7 cells treated alone. In addition, the MCF10A normal mammary cell line was used as a positive control. Western blotting was performed to confirm the overexpression of ING4 protein in the treatment groups. Unlike in the control groups, the overexpression of ING4 was clear in the wells of the treatment group.
Conclusion:
Our findings suggest that ING4 gene delivery using prepared PEI-based nonviral delivery systems is a promising approach for breast cancer treatment.
Insights
Polyethyleneimine-based polyplexes effectively delivered the inhibitor of growth 4 (ING4) gene to breast cancer cells, significantly reducing cell viability. This non-viral gene delivery system shows promise for breast cancer therapy.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Oncology
Background:
- Breast cancer is a leading cause of cancer death in women.
- Inhibitor of growth (ING) gene levels are often reduced in cancer cells.
- ING4 presents a potential therapeutic target for breast cancer.
Purpose of the Study:
- To formulate and characterize a polyplex for delivering the pING4 gene to breast cancer cells.
- To evaluate the anticancer efficacy of pING4 gene delivery.
- To assess the safety and effectiveness of the non-viral delivery system.
Main Methods:
- Synthesis and characterization of polyethyleneimine (PEI)-based non-viral vectors.
- Complexation of plasmid DNA (pING4) with PEI vectors via electrostatic interactions.
- In vitro evaluation of polyplexes on MCF-7 breast cancer cells using XTT assay and Western blot analysis.
Main Results:
- Polyplexes carrying the ING4 gene significantly reduced breast cancer cell viability compared to controls (p=0.0067).
- Maximum reduction in cell viability was observed on day 4 of the 5-day assay.
- Western blot confirmed successful overexpression of ING4 protein in treated cells, with no observed toxicity to MCF-7 or MCF10A cells.
Conclusions:
- PEI-based non-viral delivery systems are effective for ING4 gene delivery to breast cancer cells.
- ING4 gene therapy holds promise as a potential treatment strategy for breast cancer.
- Further research into this non-viral gene delivery approach is warranted.
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