Development of Dual Functional Nucleic Acid Delivery Nanosystem for DNA Induced Silencing of Bcl-2 Oncogene

Somayeh Karimi,1,* Mohamad Hassan Fouani,2,* Ahmad Moshaii,1 Maryam Nikkhah,2 Saman Hosseinkhani,3 Reza Sheikhnejad4 1Department of Physics, Tarbiat Modares University, Tehran, Iran; 2Department of Nanobiotechnology, Tarbiat Modares University, Tehran, Iran; 3Department of Biochemistry, Tarbiat Modares University, Tehran, Iran; 4Department of Molecular Biology, Tofigh Daru Engineering-Research Co., Tehran, Iran*These authors contributed equally to this workCorrespondence: Ahmad Moshaii; Maryam Nikkhah Tel +98 21 82 88 34 59; +98 21 82 88 47 34Email moshaii@modares.ac.ir; m_nikkhah@modares.ac.irIntroduction: Cancer treatment using functionalized vehicles in order to block involved genes has attracted a remarkable interest. In this study, we investigated the cellular uptake and cytotoxic effects of three sizes of anti Bcl-2 DNAi-conjugated gold nanoparticles by MCF-7 cells.Methods: Three different sizes of gold nanoparticles were synthesized by citrate reduction method and after characterization, the nanoparticles were functionalized by Bcl-2 targeted DNAi. Cell internalization of the nanoparticles was analyzed by atomic absorption spectroscopy and light microscopy. The cytotoxic effects of the nanoparticles were investigated by MTT assay, flow cytometry and RT-PCR of the target gene.Results: While poor cell internalization of bare gold nanoparticles was observed, the results demonstrated that cellular uptake of DNAi-conjugated gold nanoparticles is completely size-dependent, and the largest nanoparticle (∼ 42 nm) revealed the highest internalization rate compared to other sizes (∼ 14 and ∼ 26 nm). Experimental findings showed that the DNAi-conjugated gold nanoparticles induced apoptotic pathway by silencing of the targeted Bcl-2 gene. In addition, supplementary theoretical studies demonstrated that the 42 nm DNAi-conjugated gold nanoparticles have great photothermal conversion efficiency for treatment under external illumination and these nanoparticles can be induced further cytotoxic effect by approximately 10°C temperature elevations.Conclusion: Remarkable photothermal properties of DNAi-conjugated 42 nm Au-NPs in parallel with their high cell internalization and cytotoxic effects introduce them as potential dual functional anticancer nanosystems.Keywords: breast cancer, apoptosis, DNAi, Bcl-2, gold nanoparticles