RNAi-based therapeutic nanostrategy: IL-8 gene silencing in pancreatic cancer cells using gold nanorods delivery vehicles

• Published in: Nanotechnology Vol. 26; no. 36; pp. 365101 - 14
• Main Authors: Panwar, Nishtha, Yang, Chengbin, Yin, Feng, Yoon, Ho Sup, Chuan, Tjin Swee, Yong, Ken-Tye
• Format: Journal Article
• Language: English
• Published: England IOP Publishing 11.09.2015
• Subjects: Apoptosis; Cancer; cancer therapy; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Gene expression; Genes; Gold; Gold - chemistry; gold nanorods; Humans; Interleukin-8 - antagonists & inhibitors; Interleukin-8 - genetics; Mutations; Nanorods; Nanostructure; Nanotubes - chemistry; pancreatic cancer; Pancreatic Neoplasms - genetics; Pancreatic Neoplasms - immunology; Pancreatic Neoplasms - therapy; Ribonucleic acids; RNA, Small Interfering - pharmacology; RNAi Therapeutics - methods; siRNA
• ISSN: 0957-4484; 1361-6528
• DOI: 10.1088/0957-4484/26/36/365101

RNA interference (RNAi)-based gene silencing possesses great ability for therapeutic intervention in pancreatic cancer. Among various oncogene mutations, Interleukin-8 (IL-8) gene mutations are found to be overexpressed in many pancreatic cell lines. In this work, we demonstrate IL-8 gene silencing by employing an RNAi-based gene therapy approach and this is achieved by using gold nanorods (AuNRs) for efficient delivery of IL-8 small interfering RNA (siRNA) to the pancreatic cell lines of MiaPaCa-2 and Panc-1. Upon comparing to Panc-1 cells, we found that the dominant expression of the IL-8 gene in MiaPaCa-2 cells resulted in an aggressive behavior towards the processes of cell invasion and metastasis. We have hence investigated the suitability of using AuNRs as novel non-viral nanocarriers for the efficient uptake and delivery of IL-8 siRNA in realizing gene knockdown of both MiaPaCa-2 and Panc-1 cells. Flow cytometry and fluorescence imaging techniques have been applied to confirm transfection and release of IL-8 siRNA. The ratio of AuNRs and siRNA has been optimized and transfection efficiencies as high as 88.40 2.14% have been achieved. Upon successful delivery of IL-8 siRNA into cancer cells, the effects of IL-8 gene knockdown are quantified in terms of gene expression, cell invasion, cell migration and cell apoptosis assays. Statistical comparative studies for both MiaPaCa-2 and Panc-1 cells are presented in this work. IL-8 gene silencing has been demonstrated with knockdown efficiencies of 81.02 10.14% and 75.73 6.41% in MiaPaCa-2 and Panc-1 cells, respectively. Our results are then compared with a commercial transfection reagent, Oligofectamine, serving as positive control. The gene knockdown results illustrate the potential role of AuNRs as non-viral gene delivery vehicles for RNAi-based targeted cancer therapy applications.