title: Targeted microbubbles for ultrasound mediated gene transfection and apoptosis induction in ovarian cancer cells.
abstract: Ultrasound-targeted microbubble destruction (UTMD) technique can be potentially used for non-viral delivery of gene therapy. Targeting wild-type p53 (wtp53) tumor suppressor gene may provide a clinically promising treatment for patients with ovarian cancer. However, UTMD mediated gene therapy typically uses non-targeted microbubbles with suboptimal gene transfection efficiency. We synthesized a targeted microbubble agent for UTMD mediated wtp53 gene therapy in ovarian cancer cells. Lipid microbubbles were conjugated with a Luteinizing Hormone-Releasing Hormone analog (LHRHa) via an avidin-biotin linkage to target the ovarian cancer A2780/DDP cells that express LHRH receptors. The microbubbles were mixed with the pEGFP-N1-wtp53 plasmid. Upon exposure to 1 MHz pulsed ultrasound beam (0.5 W/cm(2)) for 30s, the wtp53 gene was transfected to the ovarian cancer cells. The transfection efficiency was (43.90 ± 6.19)%. The expression of wtp53 mRNA after transfection was (97.08 ± 12.18)%. The cell apoptosis rate after gene therapy was (39.67 ± 5.95)%. In comparison with the other treatment groups, ultrasound mediation of targeted microbubbles yielded higher transfection efficiency and higher cell apoptosis rate (p<0.05). Our experiment verifies the hypothesis that ultrasound mediation of targeted microbubbles will enhance the gene transfection efficiency in ovarian cancer cells.
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Shufang
authors: S
authors: Chang
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Juan
authors: J
authors: Guo
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Jiangchuan
authors: J
authors: Sun
authors: Department of Pharmacy, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors: Shenyin
authors: S
authors: Zhu
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Yu
authors: Y
authors: Yan
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Yi
authors: Y
authors: Zhu
authors: Department of Obstetrics and Gynecology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Min
authors: M
authors: Li
authors: Institute of Ultrasound Imaging, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
authors: Zhigang
authors: Z
authors: Wang
authors: Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
authors: Ronald X
authors: RX
authors: Xu
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D006801:Humans; D045423:Microbubbles; D010051:Ovarian Neoplasms / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011966:Receptors, LHRH / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D014465:Ultrasonics / Q000379:methods*
chemical_list: D012333:RNA, Messenger; D011966:Receptors, LHRH; D016159:Tumor Suppressor Protein p53; D007987:Gonadotropin-Releasing Hormone