Lipid nanoparticle-mediated co-delivery of siRNA and paclitaxel for efficient SLC3A2 gene silencing in pancreatic cancer cells

Abstract

Paclitaxel, a widely used anticancer drug, stabilizes microtubules and induces apoptosis; however, its efficacy is often limited by drug resistance and poor tumor specificity. To overcome these limitations, we developed a lipid nanoparticle (LNPs) for the co-delivery of siRNA targeting the SLC3A2 gene and paclitaxel. LNPs were formulated using DOTAP, DSPC, cholesterol, and PEG2000-DSPE in a molar ratio of 40:10:38.5:1.5. These lipid mixtures were combined with aqueous siRNA solutions at volume ratios of 1:5 and 1:10. Paclitaxel (25 µM) was subsequently incorporated via an ethanolic solution, and encapsulation efficiency was evaluated. The resulting paclitaxel-LNP-siRNA complexes were transfected into Mia PaCa-2 pancreatic cancer cells. Gene silencing efficacy was assessed using qPCR, western blotting, and confocal laser scanning microscopy. The paclitaxel-LNP-siRNA complexes exhibited a high encapsulation efficiency of 96.42% and demonstrated stability for over 30 days. qPCR analysis revealed a 93% reduction in SLC3A2 gene expression, while western blotting showed an 82.3% decrease in protein levels. Confocal imaging confirmed successful transfection and gene silencing, evidenced by reduced fluorescence intensity in treated cells. These findings highlight the promise of a dual-delivery system aimed at enhancing anticancer effects through the combination of gene silencing and chemotherapy in pancreatic ductal adenocarcinoma.

KEYWORDS:

Gene Silencing, Nanoparticle-based Drug Delivery, Paclitaxel, Pancreatic Ductal Adenocarcinoma, Small Interfering RNA