Combination blockage of CD73 and gp130 in cancer cells by siRNA-loaded nanoparticles

Abstract

Tumor inhibitory microenvironment plays a decisive role in tumor growth and development as well as the effectiveness of various cancer immunotherapy methods. A variety of factors in the cancer microenvironment cause these effects, including adenosine and IL-6. These two factors are among the most important goals of cancer immunotherapy by helping cancer cells to multiply and proliferate as well as inhibit antitumor immune responses. The CD73 molecule, the main enzyme expressed on the surface of many cancer cells and plays an important role in the production of adenosine in the tumor environment. The gp130 molecule is also the major cytokine IL-6 receptor in the tumor environment. Therefore, in this study, we decided to inhibit the expression of CD73 and gp130 molecules in cancer cells in combination. Methods: In this study, trimethymethyl chitosan-alginate nanoparticles loaded with siRNA molecules were used to inhibit the expression of CD73 and gp130 molecules. Physicochemical properties of nanoparticles synthesized by DLS were evaluated. The chemical structure of the synthesized nanoparticles was also investigated using H1NMR and FTIR. SEM electron microscopy was used to study the morphology of nanoparticles and electrophoresis was used to evaluate the loading power of nanoparticles and serum stability. The lethal effects of nanoparticles loaded with siRNA molecules were determined using MTT assay. The entry of nanoparticles into the cells was also examined by confocal microscopy. The expression of target genes in cancer cells was assessed by qRT-PCR test. Results: Analysis of the results of physicochemical evaluation of nanoparticles showed that they have a size of about ~ 110 nm, with a PDI <0.2 and a zeta potential of 18 mV. The nanoparticles had a spherical morphology and a high capacity for siRNA loading. The nanoparticles were also highly stable in serum and exerted little toxicity on cells. Combination therapy also had a significant effect on reducing the expression of genes involved in tumor progression, indicating the anti-tumor effect of this treatment.