Silencing of HMGA2 by siRNA Loaded Methotrexate Functionalized Polyamidoamine Dendrimer for Human Breast Cancer Cell Therapy.

Fereydoon Abedi Gaballu,Jafar Ezzati Nazhad Dolatabadi,Behzad Mansoori,Soheil Abbaspour-Ravasjani,Ali Mohammadi,Amir Zarebkohan,Ayuob Aghanejad,Gholamreza Dehghan,William Chi-Shing Cho,Nader Sheibani,Behzad Baradaran

doi:10.3390/genes12071102

Abstract

The transcription factor high mobility group protein A2 (HMGA2) plays an important role in the pathogenesis of some cancers including breast cancer. Polyamidoamine dendrimer generation 4 is a kind of highly branched polymeric nanoparticle with surface charge and highest density peripheral groups that allow ligands or therapeutic agents to attach it, thereby facilitating target delivery. Here, methotrexate (MTX)- modified polyamidoamine dendrimer generation 4 (G4) (G4/MTX) was generated to deliver specific small interface RNA (siRNA) for suppressing HMGA2 expression and the consequent effects on folate receptor (FR) expressing human breast cancer cell lines (MCF-7, MDA-MB-231). We observed that HMGA2 siRNA was electrostatically adsorbed on the surface of the G4/MTX nanocarrier for constructing a G4/MTX-siRNA nano-complex which was verified by changing the final particle size and zeta potential. The release of MTX and siRNA from synthesized nanocomplexes was found in a time- and pH-dependent manner. We know that MTX targets FR. Interestingly, G4/MTX-siRNA demonstrates significant cellular internalization and gene silencing efficacy when compared to the control. Besides, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay demonstrated selective cell cytotoxicity depending on the FR expressing in a dose-dependent manner. The gene silencing and protein downregulation of HMGA2 by G4/MTX-siRNA was observed and could significantly induce cell apoptosis in MCF-7 and MDA-MB-231 cancer cells compared to the control group. Based on the findings, we suggest that the newly developed G4/MTX-siRNA nano-complex may be a promising strategy to increase apoptosis induction through HMGA2 suppression as a therapeutic target in human breast cancer.

Highlights

IntroductionCancer is the second most common cause of death after cardiovascular disease [1]
Cancer is the second most common cause of death after cardiovascular disease [1].Surgery, chemotherapy, radiotherapy, targeted therapy and their combination are employed to manage various types of cancers in clinics [2]
The possible interaction between generation 4 (G4) dendrimer and MTX was determined by Fourier-transform infrared (FTIR) spectroscopy

Summary

Introduction

Cancer is the second most common cause of death after cardiovascular disease [1]. Chemotherapy, radiotherapy, targeted therapy and their combination are employed to manage various types of cancers in clinics [2]. Chemotherapy is the most common option to treat cancers. Following the rapid progression of nanotechnology, drugs incorporating nanoparticles have been examined to prolong blood circulation time, increase therapeutic tumor accumulation, promote anti-cancer activity, reduce undesirable side effects, and overcome multidrug resistance [3,4]. Several drug delivery systems with multiple mechanistic functions have been reported [5]. Polymers, dendrimers, and lipid-based delivery systems have demonstrated encouraging therapeutic activities in both in vitro and in vivo conditions [6]

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