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Survivin Small Interfering RNA Transfected With a Microbubble and Ultrasound Exposure Inducing Apoptosis in Ovarian Carcinoma Cells
  1. Jian jun Wang, MD*,
  2. Ying Zheng, MD,
  3. Fang Yang, MD*,
  4. Ping Zhao, PhD and
  5. Huai fang Li, MD*
  1. *Department of Obstetrics and Gynaecology, Tongji Hospital of Tongji University;
  2. Department of Ultrasound, Xuhui District Central Hospital; and
  3. Department of Microbiology, Second Military Medical University, Shanghai, China.
  1. Address correspondence and reprint requests to Huai fang Li, MD, Department of Obstetrics and Gynecology, Tongji Hospital, Tongji University School of Medicine, Shanghai, China 200065. E-mail: jjwang0524{at}yahoo.com.cn.

Abstract

Introduction: 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.

  • Survivin
  • RNA interference
  • Microbubble contrast agent
  • Ultrasound exposure
  • Ovarian carcinoma cell
  • Apoptosis

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