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Published on: May 1, 2018
In vitro evaluation of optimized liposomes for delivery of small interfering RNA
Tianzhi Yang1, Tracy Bantegui, Kaitlynn Pike
1Department of Basic Pharmaceutical Sciences, School of Pharmacy, Husson University , Bangor, ME , USA.
Abstract:
One of the biggest challenges for small interfering RNAs (siRNAs) as therapeutic agents is their insufficient cellular delivery efficiency. We developed long circulating and cationic liposomes to improve the cell uptake and inhibitory effectiveness of siRNA on the expression of vascular endothelial growth factor (VEGF) in cancer cells. SiRNA liposomes were obtained by polyelectrolyte complexation between negatively charged siRNA and positively charged liposome prepared by a hydration method. Gel electrophoresis was used to evaluate the loading efficiency of siRNA on the cationic liposome. The optimized siRNA liposomes were observed to be spherical in shape and had smooth surfaces with particle sizes of 167.7 ± 2.0 nm and zeta potentials of 4.03 ± 0.69 mV, which had no significant change when stored at 4 °C for three months. Fluorescence-activated cell sorting studies and confocal laser scanning images indicated that the cationic liposomes significantly increased the uptake of fluorescence-labeled siRNA in cancer cells. Effects of the siRNA on the inhibition of VEGF were tested by measuring concentrations of VEGF in cell culture media via an enzyme-linked immunosorbent assay and intracellular VEGF levels using a western blotting method. The liposomal siRNA was significantly effective at inhibiting the expression of VEGF in lung, liver and breast cancer cells. Optimal liposomes could effectively deliver siRNA into cancer cells and inhibit VEGF as a therapy agent.
Insights
Cationic liposomes enhance cellular delivery of small interfering RNAs (siRNAs) for cancer therapy. These liposomes effectively deliver siRNA into cancer cells, inhibiting vascular endothelial growth factor (VEGF) expression and offering a promising therapeutic approach.
Area of Science:
- Biotechnology
- Nanomedicine
- Cancer Therapeutics
Background:
- Small interfering RNAs (siRNAs) face challenges in cellular delivery for therapeutic applications.
- Vascular Endothelial Growth Factor (VEGF) is a key target in cancer therapy.
Purpose of the Study:
- To develop long-circulating, cationic liposomes for improved siRNA delivery and enhanced inhibition of VEGF expression in cancer cells.
- To evaluate the physicochemical properties, cellular uptake, and therapeutic efficacy of siRNA-loaded liposomes.
Main Methods:
- siRNA was complexed with cationic liposomes prepared via a hydration method.
- Gel electrophoresis assessed siRNA loading efficiency.
- Particle size, zeta potential, and stability were characterized.
- Cellular uptake was analyzed using fluorescence-activated cell sorting and confocal microscopy.
- VEGF inhibition was quantified using ELISA and Western blotting.
Main Results:
- Optimized siRNA liposomes exhibited spherical morphology, smooth surfaces, and stable physicochemical properties (167.7 ± 2.0 nm size, 4.03 ± 0.69 mV zeta potential) over three months.
- Cationic liposomes significantly enhanced siRNA uptake in cancer cells.
- Liposomal siRNA demonstrated significant inhibition of VEGF expression in lung, liver, and breast cancer cells.
Conclusions:
- Cationic liposomes are effective carriers for siRNA delivery into cancer cells.
- This liposomal siRNA formulation shows significant potential for inhibiting VEGF expression and could serve as a therapeutic agent for various cancers.
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