title: Experimental research of RB94 gene transfection into retinoblastoma cells using ultrasound-targeted microbubble destruction.
abstract: The purpose of this study was to explore the transfection of the recombinant expression plasmid pEGFP-C1/RB94 into human retinoblastoma cells (HXO-Rb44) using ultrasound-targeted microbubble destruction (UTMD). pEGFP-C1/RB94 was transfected into HXO-Rb44 in vitro by UTMD, with liposome as the positive control. After 24 to 72 h, the expression of the reporter gene enhanced green fluorescent protein (EGFP) was observed using fluorescent microscopy and flow cytometry. The cell viability of HXO-Rb44 was measured by a MTT assay. The mRNA and proteins of RB94, caspase-3 and Bax were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Moreover, the apoptosis rate and cell cycle progression of the cells were detected by flow cytometry. This study demonstrated that UTMD can enhance the transfection efficiency of RB94, which has an obvious impact on the inhibition of the growth process of retinoblastoma cells, suggesting that the combination of UTMD and RB94 compounds might be a useful tool for use in the gene therapy of retinoblastoma.
authors.affiliation: Department of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, P R China.
authors.forename: Min-Ming
authors.initials: MM
authors.lastname: Zheng
authors.forename: Xi-Yuan
authors.initials: XY
authors.lastname: Zhou
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Wang
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Wang
mesh_terms: D000704:Analysis of Variance; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008081:Liposomes; D045423:Microbubbles; D008856:Microscopy, Fluorescence; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis; D012175:Retinoblastoma / Q000628:therapy*; D016160:Retinoblastoma Protein / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013010:Sonication / Q000379:methods*; D014162:Transfection / Q000379:methods*
chemical_list: D008081:Liposomes; D012333:RNA, Messenger; D016160:Retinoblastoma Protein; D049452:Green Fluorescent Proteins