Skip to main content

Advertisement

SpringerLink
Log in

Enhancement of survivin gene downregulation and cell apoptosis by a novel combination: liposome microbubbles and ultrasound exposure

  • Original Paper
  • Published:
Medical Oncology Aims and scope Submit manuscript

Abstract

Ultrasound-mediated microbubble destruction (sonoporation) is an efficient and safe nonviral technique for gene delivery. In the present work, we hypothesized that short hairpin RNA (shRNA) interference therapy targeting human Survivin gene could be transfected by the novel combination of ultrasound exposure (USE) and liposome microbubbles (LM). ShRNA vectors targeting Survivin were constructed and transfected under USE and LM conditions. The optimal transfection efficiency and cell injury were compared with those of polyethylenimine (PEI)-mediated transfection in different cancer cell lines (HeLa, HepG2, Ishikawa, MCF-7, and B16-F10). The effects of gene downregulation and cell apoptosis were further investigated. The results indicated that P + USE + LM group could significantly increase the gene expression as compared with plasmid group, plasmid + USE group, plasmid + LM group (P < 0.001). The transfection efficiency of the novel combination was nearly equal to PEI-mediated transfection in some cancer cell lines while the cell viability did not decrease markedly. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis also confirmed that Survivin mRNA and protein expression could be knocked down significantly by shRNA transfection under USE and LM condition (P < 0.001). This is the first study to verify the role of shRNA therapy in vitro with novel combination of USE and LM. We concluded that this nonviral technique would be valuable in the gene transfection of shRNA and Survivin gene downregulation would lead to apparent cell apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Aoi A, Watanabe Y, Mori S, et al. Herpes simplex virus thymidine kinase-mediated suicide gene therapy using nano/microbubbles and ultrasound. Ultrasound Med Biol. 2008;34(3):425–34. doi:10.1016/j.ultrasmedbio.2007.09.004.

    Article  PubMed  Google Scholar 

  2. Yoon CS, Jung HS, Kim TK, et al. Comparison of the efficiency and toxicity of sonoporation with branched polyethylenimine-mediated gene transfection in various cultured cell lines. J Drug Target. 2008;16(10):773–9. doi:10.1080/10611860802470549.

    Article  CAS  PubMed  Google Scholar 

  3. Liang HD, Lu QL, Xue SA, et al. Optimisation of ultrasound-mediated gene transfer (sonoporation) in skeletal muscle cells. Ultrasound Med Biol. 2004;30(11):1523–9. doi:10.1016/j.ultrasmedbio.2004.08.021.

    Article  PubMed  Google Scholar 

  4. Newman CM, Bettinger T. Gene therapy progress and prospects: ultrasound for gene transfer. Gene Ther. 2007;14(6):465–75. doi:10.1038/sj.gt.3302925.

    Article  CAS  PubMed  Google Scholar 

  5. Yang F, Gu N, Chen D, et al. Experimental study on cell self-sealing during sonoporation. J Control Release. 2008;131(3):205–10. doi:10.1016/j.jconrel.2008.07.038.

    Article  CAS  PubMed  Google Scholar 

  6. Shen ZP, Brayman AA, Chen L, Miao CH. Ultrasound with microbubbles enhances gene expression of plasmid DNA in the liver via intraportal delivery. Gene Ther. 2008;15(16):1147–55. doi:10.1038/gt.2008.51.

    Article  CAS  PubMed  Google Scholar 

  7. Li YS, Reid CN, McHale AP. Enhancing ultrasound-mediated cell membrane permeabilisation (sonoporation) using a high frequency pulse regime and implications for ultrasound-aided cancer chemotherapy. Cancer Lett. 2008;266(2):156–62. doi:10.1016/j.canlet.2008.02.041.

    Article  CAS  PubMed  Google Scholar 

  8. Leong-Poi H, Kuliszewski MA, Lekas M, et al. Therapeutic arteriogenesis by ultrasound-mediated VEGF165 plasmid gene delivery to chronically ischemic skeletal muscle. Circ Res. 2007;101(3):295–303. doi:10.1161/CIRCRESAHA.107.148676.

    Article  CAS  PubMed  Google Scholar 

  9. Suzuki R, Takizawa T, Negishi Y, et al. Gene delivery by combination of novel liposomal bubbles with perfluoropropane and ultrasound. J Control Release. 2007;117(1):130–6. doi:10.1016/j.jconrel.2006.09.008.

    Article  CAS  PubMed  Google Scholar 

  10. Larina IV, Evers BM, Esenaliev RO. Optimal drug and gene delivery in cancer cells by ultrasound-induced cavitation. Anticancer Res. 2005;25(1A):149–56.

    CAS  PubMed  Google Scholar 

  11. Rahim A, Taylor SL, Bush NL, et al. Physical parameters affecting ultrasound/microbubble-mediated gene delivery efficiency in vitro. Ultrasound Med Biol. 2006;32(8):1269–79. doi:10.1016/j.ultrasmedbio.2006.04.014.

    Article  PubMed  Google Scholar 

  12. Meijering BD, Henning RH, Van Gilst WH, et al. Optimization of ultrasound and microbubbles targeted gene delivery to cultured primary endothelial cells. J Drug Target. 2007;15(10):664–71. doi:10.1080/10611860701605088.

    Article  CAS  PubMed  Google Scholar 

  13. Duvshani-Eshet M, Benny O, Morgenstern A, Machluf M. Therapeutic ultrasound facilitates antiangiogenic gene delivery and inhibits prostate tumor growth. Mol Cancer Ther. 2007;6(8):2371–82. doi:10.1158/1535-7163.MCT-07-0019.

    Article  CAS  PubMed  Google Scholar 

  14. Caplen NJ. Gene therapy progress and prospects. Downregulating gene expression: the impact of RNA interference. Gene Ther. 2004;11(16):1241–8. doi:10.1038/sj.gt.3302324.

    Article  CAS  PubMed  Google Scholar 

  15. McAnuff MA, Rettig GR, Rice KG. Potency of siRNA versus shRNA mediated knockdown in vivo. J Pharm Sci. 2007;96(11):2922–30. doi:10.1002/jps.20968.

    Article  CAS  PubMed  Google Scholar 

  16. Vandenbroucke RE, Lentacker I, Demeester J, De Smedt SC, Sanders NN. Ultrasound assisted siRNA delivery using PEG-siPlex loaded microbubbles. J Control Release. 2008;126(3):265–73. doi:10.1016/j.jconrel.2007.12.001.

    Article  CAS  PubMed  Google Scholar 

  17. Negishi Y, Endo Y, Fukuyama T, et al. Delivery of siRNA into the cytoplasm by liposomal bubbles and ultrasound. J Control Release. 2008;132(2):124–30. doi:10.1016/j.jconrel.2008.08.019.

    Article  CAS  PubMed  Google Scholar 

  18. Neu M, Fischer D, Kissel T. Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives. J Gene Med. 2005;7(8):992–1009. doi:10.1002/jgm.773.

    Article  CAS  PubMed  Google Scholar 

  19. Ho J, Wang H, Forde GM. Process considerations related to the microencapsulation of plasmid DNA via ultrasonic atomization. Biotechnol Bioeng. 2008;101(1):172–81. doi:10.1002/bit.21876.

    Article  CAS  PubMed  Google Scholar 

  20. Weerasinghe P, Li Y, Guan Y, et al. T40214/PEI complex: a potent therapeutics for prostate cancer that targets STAT3 signaling. Prostate. 2008;68(13):1430–42. doi:10.1002/pros.20807.

    Article  CAS  PubMed  Google Scholar 

  21. Boussif O, Lezoualc’h F, Zanta MA, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA. 1995;92(16):7297–301. doi:10.1073/pnas.92.16.7297.

    Article  CAS  PubMed  Google Scholar 

  22. Mehier-Humbert S, Bettinger T, Yan F, Guy RH. Ultrasound-mediated gene delivery: Kinetics of plasmid internalization and gene expression. J Control Release. 2005;104(1):203–11. doi:10.1016/j.jconrel.2005.01.011.

    Article  CAS  PubMed  Google Scholar 

  23. Chen ZY, Xie MX, Wang XF, Lü Q, Ding SW. Effects of lipid shell microbubble on ultrasound mediated EGFP gene delivery to transplanted tumors: initial experience. Chinese-German J Clin Oncol. 2008;7(7):424–8. doi:10.1007/s10330-008-0058-3.

    Article  CAS  Google Scholar 

  24. Chen S, Ding JH, Bekeredjian R, et al. Efficient gene delivery to pancreatic islets with ultrasonic microbubble destruction technology. Proc Natl Acad Sci. 2006;103(22):8469–74. doi:10.1073/pnas.0602921103.

    Article  CAS  PubMed  Google Scholar 

  25. Sangfelt SH. Apoptosis and cell growth inhibition as antitumor effector functions of interferons. Med Oncol. 2001;18(1):3–14. doi:10.1385/MO:18:1:3.

    Article  CAS  PubMed  Google Scholar 

  26. Harakeh S, Abu-El-Ardat K, Diab-Assaf M, et al. Epigallocatechin-3-gallate induces apoptosis and cell cycle arrest in HTLV-1-positive and -negative leukemia cells. Med Oncol. 2008;25(1):30–9. doi:10.1007/s12032-007-0036-6.

    Article  CAS  PubMed  Google Scholar 

  27. Altieri DC. Validating survivin as a cancer therapeutic target. Nat Rev Cancer. 2003;3(1):46–54. doi:10.1038/nrc968.

    Article  CAS  PubMed  Google Scholar 

  28. Jiang G, Li J, Zeng Z, Xian L. Lentivirus-mediated gene therapy by suppressing survivin in BALB/c nude mice bearing oral squamous cell carcinoma. Cancer Biol Ther. 2006;5(4):435–40.

    Article  CAS  PubMed  Google Scholar 

  29. Caldas H, Holloway MP, Hall BM, Qualman SJ, Altura RA. Survivin-directed RNA interference cocktail is a potent suppressor of tumour growth in vivo. J Med Genet. 2006;43(2):119–28. doi:10.1136/jmg.2005.034686.

    Article  CAS  PubMed  Google Scholar 

  30. Bian AN, Gao YH, Tan KB, et al. Preparation of human hepatocellular carcinoma-targeted liposome microbubbles and their immunological properties. World J Gastroenterol. 2004;10(23):3424–7.

    CAS  PubMed  Google Scholar 

  31. Liu P, Gao YH, Tan KB, Liu Z, Zuo S. Grey scale enhancement of rabbit liver and kidney by intravenous injection of a new lipid-coated ultrasound contrast agent. World J Gastroenterol. 2004;10(16):2369–72.

    CAS  PubMed  Google Scholar 

  32. Siu T, Rohling RN, Chiao M. Power density requirement of a 4 MHz micro-ultrasonic transducer for sonodynamic therapy. Biomed Microdevices. 2008;10(1):89–97. doi:10.1007/s10544-007-9113-z.

    Article  PubMed  Google Scholar 

Download references

Author information

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, Guangzhou Research Institute of Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510150, China

    Kun Liang

  3. Department of Function Imaging and Ultrasonography, Guangzhou First Municipal People’s Hospital, Guangzhou Medical College, Guangzhou, 510180, China

    Jianhua Liu

  4. 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 & Lingyun Fang

Authors
  1. Zhiyi Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kun Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianhua Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mingxing Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xinfang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Qing Lü
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lingyun Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhiyi Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, Z., Liang, K., Liu, J. et al. Enhancement of survivin gene downregulation and cell apoptosis by a novel combination: liposome microbubbles and ultrasound exposure. Med Oncol 26, 491–500 (2009). https://doi.org/10.1007/s12032-008-9161-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12032-008-9161-0

Keywords

Search

Navigation