Effects of RNA interference-mediated knockdown of livin and survivin using monomethoxypolyethylene glycol-chitosan nanoparticles in MG-63 osteosarcoma cells

Guan,Hua‑Peng; Sun,Jian‑Zhong; Feng,Xiao‑Lei; Chen,Jin‑Shui; Chen,Fang‑Jing; Cheng,Xiao‑Fei; Liu,Xin‑Wei; Ni,Bin

doi:10.3892/mmr.2015.4709

Effects of RNA interference-mediated knockdown of livin and survivin using monomethoxypolyethylene glycol-chitosan nanoparticles in MG-63 osteosarcoma cells

Authors:
- Hua‑Peng Guan
- Jian‑Zhong Sun
- Xiao‑Lei Feng
- Jin‑Shui Chen
- Fang‑Jing Chen
- Xiao‑Fei Cheng
- Xin‑Wei Liu
- Bin Ni
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Affiliations: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China, Department of Orthopedics, People's Liberation Army 456 Hospital, Jinan, Shandong 250031, P.R. China, Department of Orthopedics, General Hospital of Shenyang Military Area Command of Chinese People's Liberation Army, Rescue Center of Severe Wound and Trauma of Chinese People's Liberation Army, Shenyang, Liaoning 110840, P.R. China
Published online on: December 23, 2015 https://doi.org/10.3892/mmr.2015.4709
Pages: 1821-1826

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Abstract

MG-63 human osteosarcoma cells were transfected with short hairpin RNA (shRNA) against livin and survivin using monomethoxypolyethylene glycol‑chitosan (mPEG‑CS) nanoparticles (NPs) as carriers, with the aim of evaluating the effect on cell proliferation and apoptosis. mPEG‑CS NPs sized ~100 nm were prepared by ionic crosslinking. mPEG‑CS‑livin shRNA, mPEG‑CS‑survivin shRNA and mPEG‑CS‑(livin shRNA + survivin shRNA) NPs were constructed by electrostatic adsorption at NP suspension/gene solution ratios of 3:1 to transfect MG‑63 cells. The expression levels of livin and survivin mRNA and protein were measured by reverse transcription‑polymerase chain reaction and western blotting, respectively. The inhibitory effects of downregulated livin and survivin expression on cell proliferation were measured using an MTT assay. The apoptosis‑inducing effects of livin and surivin knockdown were investigated using a Hoechst staining kit. All shRNA groups resulted in reduced expression of livin and survivin mRNA and protein in MG‑63 cells. The MTT assay and Hoechst staining indicated that simultaneous knockdown of livin and survivin genes inhibited the proliferation of MG‑63 cells and promoted their apoptosis, to a greater extent than knocking down either gene individually. The simultaneous interference mediated by mPEG‑CS NPs significantly reduced livin and survivin expression in MG‑63 cells, suppressed proliferation and facilitated apoptosis, to a greater extent than knockdown of either livin or survivin alone were. Thus the results indicate a synergistic effect of livin and survivin.

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