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Novel ultrasound-targeted microbubble destruction mediated short hairpin RNA plasmid transfection targeting survivin inhibits gene expression and induces apoptosis of HeLa cells

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Abstract

Survivin is an attractive target for tumor growth inhibition and represents a significant approach to anticancer therapy. RNA interference is an important tool for specifically down-regulating the expression of cellular genes. However, the efficiency of short hairpin RNA (shRNA) on the expression of survivin gene and the influence on the cell apoptosis transfected by the non-viral gene transfer system of ultrasound-targeted microbubble destruction was not explored. In this work, recombinant expression plasmid of shRNA targeting survivin gene was constructed and added to cultured cervical cancer cells followed by ultrasound exposure and SonoVue® microbubble. Expression of survivin mRNA and protein were assessed by RT-PCR and western blot analysis. Apoptosis ratio was quantified by flow cytometry marked with annexin V and 7-AAD. After transfected for 48 h, the expression of survivin mRNA and protein were (16.67 ± 2.73)% and (21.33 ± 3.55)%, respectively. The apoptosis rate was (45.41 ± 1.47)%. The differences were significant as compared with other groups (P < 0.01). In conclusion, we suggested that survivin could be regarded as an ideal anticancer target of cervical cancer. Recombinant expression plasmid of shRNA targeting survivin gene mediated by ultrasound-targeted microbubble destruction technique could effectively inhibit the expression of target gene and induce cell apoptosis. This novel method for RNA interference represents a powerful, promising non-viral technology that can be used in the tumor gene therapy and research.

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Acknowledgments

We thank Mr. Zhenhui He (Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China) for helpful technical discussion about this work, thank Mr. Zhihui Liang for flow cytometry assays (College of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China), and thank Mr. Zhijiang Liang (Department of Epidemiology and Health Statistics, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China) for statistical direction.

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Authors and Affiliations

  1. Department of Medical Ultrasound, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510150, China

    Zhiyi Chen

  2. Department of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510150, China

    Kun Liang

  3. Department of Ultrasound, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, 430022, China

    Mingxing Xie, Xinfang Wang, Qing Lü & Jing Zhang

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  1. Zhiyi Chen
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  2. Kun Liang
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  3. Mingxing Xie
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  4. Xinfang Wang
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  5. Qing Lü
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  6. Jing Zhang
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Corresponding author

Correspondence to Zhiyi Chen.

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Zhiyi Chen and Kun Liang contributed equally to this work.

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Chen, Z., Liang, K., Xie, M. et al. Novel ultrasound-targeted microbubble destruction mediated short hairpin RNA plasmid transfection targeting survivin inhibits gene expression and induces apoptosis of HeLa cells. Mol Biol Rep 36, 2059–2067 (2009). https://doi.org/10.1007/s11033-008-9417-y

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  • DOI: https://doi.org/10.1007/s11033-008-9417-y

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