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Updated: Apr 28, 2026
Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae
Published on: March 25, 2015
Comparison of VEGF gene silencing efficiencies of chitosan and protamine complexes containing shRNA
Fulden Erdem-Çakmak1, Suna Özbaş-Turan, Emine Şalva
1Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, Tıbbiye Street, 34668, Istanbul, Turkey.
Abstract:
VEGF is an angiogenic factor promoting the proliferation and migration of endothelial cells. Inhibition of VEGF by RNAi mechanism is one of the novel and the most important strategies in antiangiogenesis therapy. In this study, the tumor silencing efficiency of ternary complexes after addition of protamine to chitosan complexes containing VEGF targeting shRNA was investigated. Besides chitosan, protamine is an effective gene delivery material. Binary and ternary complexes consisting of chitosan, protamine, and shRNA were prepared to target VEGF, their morphology, size, and zeta potential of the complexes being measured. The average size of the complexes was between 173 and 284 nm and zeta potential was between +10 and 16 mV. In the ternary complexes, size decreased as the chitosan ratio increased; however, its molecular weight had no effect on the size of complexes. HeLa, HEK293, and MCF-7 cell lines were used for in vitro transfection. VEGF was assayed by ELISA. A higher silencing effect was obtained using ternary complexes. Transgene expression was increased by adding protamine to chitosan complexes. Gene inhibition values in cell lines followed the rank HEK293>HeLa>MCF-7. The addition of protamine to the chitosan/shRNA (VEGF) complexes increased the knockdown of VEGF genes in the cell lines, and no cytotoxicity was found after the complexes had been incorporated into the cells.
Insights
This study shows that adding protamine to chitosan complexes effectively silences VEGF genes in cancer cells. These novel ternary complexes offer a promising, non-toxic approach for anti-angiogenesis cancer therapy.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Vascular Endothelial Growth Factor (VEGF) drives tumor angiogenesis by promoting endothelial cell proliferation and migration.
- Inhibiting VEGF via RNA interference (RNAi) is a key strategy in anti-angiogenesis cancer therapy.
- Developing efficient and safe gene delivery systems for RNAi is crucial for therapeutic success.
Purpose of the Study:
- To investigate the tumor silencing efficiency of novel ternary complexes.
- To evaluate the impact of protamine addition to chitosan/shRNA complexes targeting VEGF.
- To assess the safety and efficacy of these complexes in various cancer cell lines.
Main Methods:
- Preparation and characterization of binary and ternary complexes of chitosan, protamine, and VEGF-targeting shRNA.
- Measurement of complex size, zeta potential, and morphology.
- In vitro transfection of HeLa, HEK293, and MCF-7 cell lines.
- Quantification of VEGF gene silencing using ELISA.
- Cytotoxicity assessment of the complexes.
Main Results:
- Ternary complexes demonstrated enhanced VEGF gene silencing compared to binary complexes.
- The addition of protamine to chitosan/shRNA complexes significantly increased transgene expression and gene inhibition.
- Gene inhibition efficacy followed the order HEK293 > HeLa > MCF-7.
- Complexes exhibited sizes between 173-284 nm and zeta potentials of +10 to +16 mV.
- No significant cytotoxicity was observed in the tested cell lines.
Conclusions:
- Protamine-enhanced chitosan/shRNA ternary complexes are effective and non-toxic for VEGF gene silencing.
- These complexes represent a promising delivery system for anti-angiogenesis cancer therapy.
- The study highlights the potential of optimizing complex composition for targeted gene therapy applications.
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