title: Magnetic gold nanoparticle-mediated small interference RNA silencing Bag-1 gene for colon cancer therapy.
abstract: Bcl-2-associated athanogene 1 (Bag-1) is a positive regulator of Bcl-2 which is an anti-apoptotic gene. Bag-1 was very slightly expressed in normal tissues, but often highly expressed in many tumor tissues, particularly in colon cancer, which can promote metastasis, poor prognosis and anti-apoptotic function of colon cancer. We prepared and evaluated magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex, a gene therapy system, which can transfect cells efficiently, for both therapeutic effect and safety in vitro mainly by electrophoretic mobility shift assays, flow cytometric analyses, cell viability assays, western blot analyses and RT-PCR (real-time) assays. Magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex was successfully transfected into LoVo colon cancer cells and the exogenous gene was expressed in the cells. Flow cytometric results showed apoptosis rate was significantly increased. In MTT assays, magnetic gold nanoparticles revealed lower cytotoxicity than Lipofectamine 2000 transfection reagents (P<0.05). Both in western blot analyses and RT-PCR assays, magnetic gold nanoparticle/Bag-1 siRNA recombinant plasmid complex transfected cells demonstrated expression of Bag-1 mRNA (P<0.05) and protein (P<0.05) was decreased. In further study, c-myc and β-catenin which are main molecules of Wnt/β‑catenin pathway were decreased when Bag-1 were silenced in nanoparticle plasmid complex transfected LoVo cells. These results suggest that magnetic gold nanoparticle mediated siRNA silencing Bag-1 is an effective gene therapy method for colon cancer.
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China.
authors.forename: Wenbai
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China.
authors.forename: Zhan'ao
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China.
authors.forename: Guanzhou
authors.initials: G
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China.
authors.forename: Ailing
authors.initials: A
authors.lastname: Tian
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 200012, P.R. China.
authors.forename: Nianfeng
authors.initials: N
authors.lastname: Sun
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology; D017209:Apoptosis; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000473:pathology / Q000628:therapy*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D015536:Down-Regulation; D004354:Drug Screening Assays, Antitumor; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D017979:Gold Colloid / Q000008:administration & dosage* / Q000633:toxicity; D006801:Humans; D008055:Lipids / Q000008:administration & dosage / Q000633:toxicity; D059346:Magnets; D053758:Nanoparticles / Q000008:administration & dosage* / Q000633:toxicity; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: C091732:BCL2-associated athanogene 1 protein; D004268:DNA-Binding Proteins; D017979:Gold Colloid; D008055:Lipids; C086724:Lipofectamine; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; D014157:Transcription Factors