title: Survivin small interfering RNA transfected with a microbubble and ultrasound exposure inducing apoptosis in ovarian carcinoma cells.
abstract: BACKGROUND
  Small interfering RNA (siRNA) has been used to knock down the expression of specific genes. However, its delivery remains a challenge. Recently, ultrasound combined with microbubbles has been used to deliver plasmid into cells and has shown much superiority. Whether a survivin siRNA transfected with a microbubble contrast agent combined with ultrasound exposure could inhibit survivin expression and induce ovarian cancer cell apoptosis was investigated.
  
  
  METHODS
  The survivin gene was amplified and inserted into vector pEGFP-N1, resulting in psurvivin-EGFP. Three siRNA expression cassettes (SECs) targeting survivin were obtained by 2-step polymerase chain reaction. The SECs and psurvivin-EGFP were cotransfected into 293T cells. A functional SEC was selected and inserted into pMD18T, resulting in the survivin-targeting siRNA expression plasmid, which was transfected into SKOV-3 ovarian carcinoma cells using SonoVue, a microbubble contrast agent, and ultrasound exposure. Survivin expression was monitored by Western blot, and apoptosis was determined by fluorescence-activated cell sorting.
  
  
  RESULTS
  One siRNA effectively inhibited survivin expression. SonoVue with ultrasound effectively delivered survivin siRNA to SKOV-3 cells, inhibited survivin expression, and induced apoptosis.
  
  
  CONCLUSIONS
  Delivery of survivin siRNA using a microbubble contrast agent combined with ultrasound exposure can effectively inhibit survivin expression and induce apoptosis, providing a new promising approach for siRNA delivery in vivo.
authors.affiliation: Department of Obstetrics and Gynaecology, Tongji Hospital of Tongji University, Shanghai, China.
authors.forename: Jian jun
authors.initials: Jj
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.forename: Huai fang
authors.initials: Hf
authors.lastname: Li
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D045423:Microbubbles; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D014162:Transfection; D014465:Ultrasonics
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000077022:Survivin