In vitro gene silencing effect of chitosan/shRNA PDGF-D nanoparticles in breast cancer

Abstract

Breast cancer is the most common cancer worldwide in women

and it is highly malignant and fatal. PDGF-D plays role in

regulation of many cellular processes such as angiogenesis.

PDGF-D is overexpressed in many types of cancers and promote

tumor growth and metastasis. Silencing of PDGF-D gene by

using shRNA with an appropriate carrier system may decrease

tumor growth and metastasis. In our study, we prepared chitosan

nanoparticles loaded with five different shRNA plasmids

targeting different exons of PDGF-D gene. Then, nanoparticles

were characterized in vitro and transfection efficiency of these

nanoparticles were investigated in breast cancer cell lines (MCF-

7, MDA-MB-231 and MDA-MB-435). The effects of single

and multiple shRNA sequences, molecular weight of chitosan

(150 kDa and 400 kDa) and the amount of shRNA (100 and

500 μg) on the characterization and transfection efficiencies of

nanoparticles have been studied. Size of nanoparticles changed

between 200-400 nm and approximately 95-100% encapsulation

efficiency were obtained. Release of shRNA changed with the

molecular weight of chitosan. It was obtained that formulation

containing shRNA plasmid targeting PDGF-D exon 6 (NP1)

has the highest silencing efficiency in MDA-MB-231 cell line. It

was also evaluated that chitosan can be a suitable gene delivery

system for shRNA targeting PDGF-D.

Keywords

• Breast cancer,chitosan,shRNA,gene silencing,RNAi

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