Folate receptor-targeted nanoparticle delivery of HuR-RNAi suppresses lung cancer cell proliferation and migration

Journal of Nanobiotechnology

Research

Uday B. Kompella¹  Allshine Chen²  Yan Daniel Zhao³  Anish Babu⁴  Ranganayaki Muralidharan⁴  Narsireddy Amreddy⁴  Rajagopal Ramesh⁵  Anupama Munshi⁶  Kanthesh Basalingappa⁶  Meghna Mehta⁶ 

[1] Department of Pharmaceutical Sciences and Opthalmology, University of Colorado, 80045, Denver, CO, USA;Departments of Epidemiology and Statistics, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Departments of Epidemiology and Statistics, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Stephenson Cancer Center, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Departments of Pathology, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Stephenson Cancer Center, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Departments of Pathology, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Stephenson Cancer Center, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Graduate Program in Biomedical Sciences, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Department of Pathology, Stanton L. Young Biomedical Research Center, Suite 1403, 975 N.E., 10th Street, 73104, Oklahoma City, OK, USA;Departments of Radiation Oncology, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;Stephenson Cancer Center, University of Oklahoma Health Sciences Center, 73104, Oklahoma City, OK, USA;

关键词: Folate;    HuR;    Lung cancer;    Nanoparticle;    Oncoprotein;    siRNA;    Targeting;   

DOI  :  10.1186/s12951-016-0201-1

 received in 2016-04-12, accepted in 2016-06-02,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundHuman antigen R (HuR) is an RNA binding protein that is overexpressed in many human cancers, including lung cancer, and has been shown to regulate the expression of several oncoproteins. Further, HuR overexpression in cancer cells has been associated with poor-prognosis and therapy resistance. Therefore, we hypothesized that targeted inhibition of HuR in cancer cells should suppress several HuR-regulated oncoproteins resulting in an effective anticancer efficacy. To test our hypothesis, in the present study we investigated the efficacy of folate receptor-α (FRA)-targeted DOTAP:Cholesterol lipid nanoparticles carrying HuR siRNA (HuR-FNP) against human lung cancer cells.ResultsThe therapeutic efficacy of HuR-FNP was tested in FRA overexpressing human H1299 lung cancer cell line and compared to normal lung fibroblast (CCD16) cells that had low to no FRA expression. Physico-chemical characterization studies showed HuR-FNP particle size was 303.3 nm in diameter and had a positive surface charge (+4.3 mV). Gel retardation and serum stability assays showed that the FNPs were efficiently protected siRNA from rapid degradation. FNP uptake was significantly higher in H1299 cells compared to CCD16 cells indicating a receptor-dose effect. The results of competitive inhibition studies in H1299 cells demonstrated that HuR-FNPs were efficiently internalized via FRA-mediated endocytosis. Biologic studies demonstrated HuR-FNP but not C-FNP (control siRNA) induced G1 phase cell-cycle arrest and apoptosis in H1299 cells resulting in significant growth inhibition. Further, HuR-FNP exhibited significantly higher cytotoxicity against H1299 cells than it did against CCD16 cells. The reduction in H1299 cell viability was correlated with a marked decrease in HuR mRNA and protein expression. Further, reduced expression of HuR-regulated oncoproteins (cyclin D1, cyclin E, and Bcl-2) and increased p27 tumor suppressor protein were observed in HuR-FNP-treated H1299 cells but not in C-FNP-treated cells. Finally, cell migration was significantly inhibited in HuR-FNP-treated H1299 cells compared to C-FNP.ConclusionsOur results demonstrate that HuR is a molecular target for lung cancer therapy and its suppression using HuR-FNP produced significant therapeutic efficacy in vitro.

【 授权许可】

CC BY   
© The Author(s) 2016