Abstract
Surface functionalization of nanoparticles (NPs) for therapeutic siRNA delivery into cancer cells has gained interest. The present study was designed for surface functionalization of gold nanoparticles (AuNPs) for efficient siRNA delivery and knockdown in cancer cells. In order to achieve this objective, AuNPs were coated with HER2-siRNA in the presence of 11-mercaptoundecanoic acid (11-MUA), calcium chloride (CaCl2) and polyethyleneimine (PEI) in alternate charge bearing successive layers. MCF-7 cells were cultured and transfected with fabricated assembly of AuNPs. Cytotoxicity analysis revealed that the half inhibitory concentration (IC50) for the formulation was 45.35 nM . Total RNA was isolated from transfected cells, reverse transcribed into complementary DNA (cDNA) and real-time polymerase chain reaction (RT-PCR) was performed. The RT-PCR based delta-delta Ct analysis in treated cells revealed a significant 18.94 times decrease (p<0.001) in the expression of HER2 gene standardized with ACTB housekeeping gene as compared to untreated cells, which makes this formulation a potent approach for siRNA delivery and gene knockout.
Highlights
Cancer is one of the major causes of death worldwide
RNA interference (RNAi) mediated gene silencing has appeared as a powerful strategy in handling cancer producing genes by complementary base-pairing mechanism [27]. small interfering RNA (siRNA) is a very important tool with substantial use in cancer therapy [28]
Designed siRNA has the ability to modify the expression of cancer-causing genes by homology-based pairing and post-transcriptional silencing. siRNA is a powerful tool to knockdown the oncogenes due to its specific and efficient silencing of target mRNAs [29]
Summary
Cancer is one of the major causes of death worldwide. Globally, every 1 in 6 deaths is related to cancer and the number is still expected to increase [1]. Advantages of using AuNPs include their synthesis in required size with narrow size distribution range, mono dispersity, surface modification for creation of multifunctional monolayers to allow multiple moieties such as nucleic acids and targeting agents to be attached [13] They impart little cytotoxicity and efficient biodistribution in targeted cells [14,15]. Calcium related salts have been extensively studied as non-viral vectors for gene delivery, due to their inherent use as a compositional part of bones and teeth [18] They have excellent biocompatibility and play roles in cellular uptake mechanisms via endocytosis [19]. We demonstrate the successful delivery of siRNA through AuNPs and calcium-based formulation assembly for knocking down the expression of HER2 gene
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