title: Targeted Microbubbles for Ultrasound Mediated Short Hairpin RNA Plasmid Transfection to Inhibit Survivin Gene Expression and Induce Apoptosis of Ovarian Cancer A2780/DDP Cells.
abstract: Nonviral gene transfer by ultrasound-targeted microbubble destruction (UTMD) is an promising technique for RNA interference (RNAi) therapy. Targeting silence survivin gene may provide an important therapeutic option for patients with ovarian cancer. However, UTMD mediated RNAi therapy typically uses nontargeted microbubbles with suboptimal gene transfection efficiency. In this work, a LHRHa targeted microbubble agent and recombinant expression plasmid of shRNA targeting survivin gene (pshRNA survivin) were constructed for UTMD mediated pshRNA survivin therapy in ovarian cancer A2780/DDP cells that express LHRH receptors. The targeted microbubbles (TMBs) mixed with the pshRNA survivin were added to cultured ovarian cancer cells followed by ultrasound exposure (1 MHz, 0.5 W/cm(2)) for 30 s. After transfection for 48 h, the expression of survivin mRNA and protein were (0.36 ± 0.036) and (0.05 ± 0.02), respectively. The cell proliferation inhibitory rates at 24, 48, and 72 h after treatment are (42.08 ± 3.20)%, (54.60 ± 1.02)%, and (74.25 ± 2.14)%, respectively, and the apoptosis rate was (28.99 ± 2.70)%. The expression of apoptosis related protein caspase-9 and caspase-3 were (0.95 ± 0.09) and (2.6 ± 0.21). In comparison with the other treatment groups, ultrasound mediation of targeted microbubbles yielded higher RNAi efficiency and higher cell apoptosis rate and cell proliferation inhibitory rate (p < 0.05). Our experiment verifies the hypothesis that ultrasound mediation of targeted microbubbles will enhance RNAi efficiency in ovarian cancer cells. This novel method for RNA interference represents a powerful, promising no viral technology that can be used in the tumor gene therapy and research.
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.|National Engineering Research Center of Ultrasound Medicine , Chongqing 400010, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Shufang
authors.initials: S
authors.lastname: Chang
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Jiangchuan
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Pharmacy, First Affiliated Hospital of Chongqing Medical University , Chongqing 400016, China.
authors.forename: Shenyin
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Caixiu
authors.initials: C
authors.lastname: Pu
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Yaowei
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Institute of Ultrasound Imaging, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010, China.|Department of Biomedical Engineering, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Ronald X
authors.initials: RX
authors.lastname: Xu
mesh_terms: D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D049109:Cell Proliferation; D005260:Female; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D045423:Microbubbles; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D000077022:Survivin; D014162:Transfection; D014407:Tumor Cells, Cultured; D014465:Ultrasonics
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D034741:RNA, Small Interfering; D000077022:Survivin; D053148:Caspase 3; D053453:Caspase 9
keywords: RNA interference; apoptosis; microbubbles; ovarian cancer; survivin; ultrasound