Development of a carrier system containing hyaluronic acid and protamine for siRNA delivery in the treatment of melanoma

Invest New Drugs. 2021 Feb;39(1):66-76. doi: 10.1007/s10637-020-00986-3. Epub 2020 Aug 13.

Abstract

The use of small interfering RNA (siRNA) in melanoma treatment remains limited owing to its biological properties. Herein, we developed a carrier system containing hyaluronic acid and protamine for siRNA delivery. Considering zeta potential and particle size as standards, the ratio of each component in liposome nanoparticles prepared was screened using the control variable method, and siRNA cationic liposome nanoparticles were prepared based on the optimal results obtained. The encapsulation rate of the cationic liposome nanoparticles was measured, and particle morphology was observed. B16F10 cells were treated with the nanoparticles; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, cell scratch experiments, and cell uptake experiments were performed to determine the effectiveness of the loaded siRNA. A mouse model was then established, and tumour tissues were subjected to haematoxylin-eosin staining. The inhibition of the survivin gene and protein expression were assessed using reverse transcription-polymerase chain reaction and western blotting, respectively. The results showed that the optimal mass ratio of hyaluronic acid (HA)-siRNA-to-protamine was 1.0; in the HA-siRNA-protamine complex containing 25 μg siRNA, the addition of 50 μL liposomes yielded optimal particles. And encapsulation rate was 85.07%. The nanoparticles demonstrated a significant inhibitory effect against melanoma cells; siRNA liposomes may inhibit tumour growth by down-regulating survivin. Survivin-siRNA cationic liposome nanoparticles could effectively inhibit the proliferation and migration of melanoma B16F10 cells in vitro and the proliferation of subcutaneous melanoma B16F10 cells, probably by inhibiting survivin mRNA and protein expression. Graphical abstract.

Keywords: Cancer therapy; Cationic liposome; Melanoma; Nanoparticle; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Chemistry, Pharmaceutical
  • Dose-Response Relationship, Drug
  • Drug Carriers / chemistry
  • Drug Stability
  • Female
  • Hyaluronic Acid / chemistry*
  • Liposomes / chemistry
  • Melanoma / pathology*
  • Mice
  • Mice, Inbred C57BL
  • Nanoparticles / chemistry*
  • Particle Size
  • RNA, Small Interfering / administration & dosage*
  • RNA, Small Interfering / pharmacology*
  • Random Allocation
  • Surface Properties
  • Survivin / drug effects

Substances

  • Drug Carriers
  • Liposomes
  • RNA, Small Interfering
  • Survivin
  • Hyaluronic Acid