Design and anti-tumor activity of self-loaded nanocarriers of siRNA.

Polypeptide carriers have a good cell compatibility, rich functionality, and facile synthesis and modification, make them promising materials as siRNA vectors. Phenylalanine dipeptide (FF) has been previously assessed as an siRNA vector and showed to have two major drawbacks, namely poor water solubility and poor serum stability. Herein, the FF backbone was modified by ligating a PEG-Arg-Ala (PEG-RA) sequence at the N-terminus to increase its hydrophilicity and serum stability. Arg is a typical amino acid in the cell penetrating peptide, which can increase the efficiency of cell internalization. Ala acts as a spacer to avoid steric hindrance. The target sequence PEG-RAFF was synthesized by a solid phase peptide synthesis. The morphology, particle size, and siRNA ratio were assessed by SEM, TEM, DLS, and gel electrophoresis. Further, MCF-7 cells were used as a model and survivin-siRNA as a passenger to assess cell internalization, inhibition of gene expression rate, and apoptosis rate using confocal microscopy, real-time PCR, and flow cytometry. At a concentration of 1 mg/mL, PEG-RAFF took the form of nanovesicles with a diameter of 154.74 ± 14.36 nm. The optimal PEG-RAFF to siRNA ratio was N/P = 100:1. Compared with the control group, the red fluorescence of TAMRA(Carboxytetramethylrhodamine, Red fluorescence)-siRNA transfected into cells was clearly visible in the confocal microscope image. The inhibition rate of survivin was 67.99 ± 10.31%, and the apoptotic rate was 16.07%. Therefore, PEG-RAFF has potential as an siRNA carrier in cancer treatment.