title: Effects of RNA interference-mediated knockdown of livin and survivin using monomethoxypolyethylene glycol-chitosan nanoparticles in MG-63 osteosarcoma cells.
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.
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Hua-Peng
authors.initials: HP
authors.lastname: Guan
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Jian-Zhong
authors.initials: JZ
authors.lastname: Sun
authors.affiliation: Department of Orthopedics, People's Liberation Army 456 Hospital, Jinan, Shandong 250031, P.R. China.
authors.forename: Xiao-Lei
authors.initials: XL
authors.lastname: Feng
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Jin-Shui
authors.initials: JS
authors.lastname: Chen
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Fang-Jing
authors.initials: FJ
authors.lastname: Chen
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Xiao-Fei
authors.initials: XF
authors.lastname: Cheng
authors.affiliation: 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.
authors.forename: Xin-Wei
authors.initials: XW
authors.lastname: Liu
authors.affiliation: Department of Spine Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Ni
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D048430:Cell Shape; D048271:Chitosan / Q000737:chemistry*; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D009363:Neoplasm Proteins / Q000378:metabolism*; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011092:Polyethylene Glycols / Q000737:chemistry*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000077022:Survivin
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C107489:BIRC5 protein, human; C419866:BIRC7 protein, human; D051034:Inhibitor of Apoptosis Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000077022:Survivin; C118638:glycol-chitosan; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol; D048271:Chitosan