Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Gene therapy for hepatocellular carcinoma using sonoporation enhanced by contrast agents

Cancer Gene Therapy volume 12, pages 884–889 (2005)Cite this article

Abstract

We examined whether sonoporation enhanced by a contrast agent (BR14) was effective in gene therapy for hepatocelluar carcinoma (HCC). Human hepatic cancer cells (SK-Hep1) and plasmid cDNAs expressing green fluorescent protein (GFP), interferonβ (IFNβ), and LacZ were used. In vitro, SK-Hep1 cell suspensions with DNA and BR14 were sonoporated. Expressions of every plasmid cDNA and the antitumor effect of IFNβ were analyzed. In vivo, GFP and IFNβ genes with BR14 were directly injected into subcutaneous tumors using SK-Hep1 in nude mice, and transcutaneous sonoporation of the tumors was performed. GFP gene transfections and tumor diameters after IFNβ gene transfection were examined. In vitro, no SK-Hep1 cells were transfected without sonication, whereas transfections were successful after sonication with BR14. Antitumor effect of IFNβ gene transfection by ultrasound (US) and with BR14 was revealed. In vivo, the SK-Hep1 cells expressed GFP, and the IFNβ gene transfection by US with BR14 reduced tumor size significantly. In conclusion, gene therapy with sonoporation enhanced by a contrast agent may become a new treatment option for HCC.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Yamashita Y, Shimada M, Tachibana K, et al. In vivo gene transfer into muscle via electrro-sonoporation. Hum Gene Ther. 2002;13:2079–2084.

    Article  CAS  PubMed  Google Scholar 

  2. Allan L, Axel FB, Sheila EF, et al. Ultrasound enhances reporter gene expression after transfection of vascular cells in vitro. Circulation. 1999;99:2617–2620.

    Article  Google Scholar 

  3. Evan CU, Thomas PM, Robert HS . Ultrasound enhances gene expression of liposomal transfection. Invest Radiol. 1997;32:723–727.

    Article  Google Scholar 

  4. Leonhard M, Srinivas S, Seung-kew Y, et al. Gene therapy of hepatocellular carcinoma in vitro and in vivo in nude mice by adenoviral transfer of the Escherichia coli purine nucleoside phosphorylase gene. Hepatology. 2000;31:606–614.

    Article  Google Scholar 

  5. Miguel B, Guillermo M, Guillem G, et al. Gene therapy of orthotopic hepatocellular carcinoma in rats using adenovirus coding for Interleukin 12. Hepatology. 2001;33:52–61.

    Article  Google Scholar 

  6. Terazaki Y, Yano S, Yuge K, et al. An optimal therapeutic expression level is crucial for suicide gene therapy for hepatic metastatic cancer in mice. Hepatology. 2003;37:155–163.

    Article  CAS  PubMed  Google Scholar 

  7. Maria TV, Andres HG, Michael I, et al. Impact of preimmunization on adenoviral vector expression and toxicity in a subcutaneous mouse cancer model. Mol Ther. 2002;6:342–348.

    Article  Google Scholar 

  8. Nakashima M, Tachibana K, Iohara K, Ito M, Ichikawa M, Akamine A . Induction of reparative dentin formation by ultrasound-mediated gene delivery of growth/differentiation factor 11. Hum Gene Ther. 2003;14:591–597.

    Article  CAS  PubMed  Google Scholar 

  9. Taniyama Y, Morishita R, Aoki M, et al. Development of safe and efficient novel nonviral gene transfer using ultrasound: enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle. Gene Therapy. 2002;9:372–380.

    Article  CAS  PubMed  Google Scholar 

  10. Victor JD, Michael JM, Ryuichi M, Yasufumi K . Fusigenic viral liposome for gene therapy in cardiovascular diseases. Proc Natl Acad Sci USA. 1996;93:11421–11425.

    Article  Google Scholar 

  11. Marshall E . Gene therapy death prompts review of adenovirus vector. Science. 1999;286:2244–2245.

    Article  CAS  PubMed  Google Scholar 

  12. Evan CU, Evan H, Mani V, Thomas M . Gene delivery using ultrasound contrast agents. Echocardiography. 2001;18:355–361.

    Article  Google Scholar 

  13. Patrick B, Valerie PC, Adolfo G, et al. Naked DNA Injection for liver metastases treatment in rats. Hepatology. 2002;35:1144–1152.

    Article  Google Scholar 

  14. Tachibana K, Uchida T, Ogawa K, Yamashita N, Tamura K . Induction of cell-membrane porosity by ultrasound. Lancet. 1999;353:1409.

    Article  CAS  PubMed  Google Scholar 

  15. Shiping B, Brian DT, Douglas LM . Transfection of a reporter plasmid into cultured cells by sonoporation in vitro. Ultrasound Med Biol. 1997;23:953–959.

    Article  Google Scholar 

  16. Shiping B, Brian DT, Richard AG, Douglas LM . In vivo transfection of melanoma cells by Lithotripter shock waves. Cancer Res. 1998;58:219–221.

    Google Scholar 

  17. Marcus F, Craig D, Robert FM, Taylor DL . Measurement of cytoplasmic pH in Dictyostelium discoideum by using a new method for introducing macromolecules into living cells. Eur J Cell Biol. 1986;40:242–247.

    Google Scholar 

  18. Ka-yun N, Yang L . Therapeutic ultrasound: its application in drug delivery. Med Res Rev. 2002;22:204–223.

    Article  Google Scholar 

  19. Mark W, Junru W, Jen-Fu C . Experimental study of optison concentration on sonoporation in vitro. Ultrasound Med Biol. 2000;26:1169–1175.

    Article  Google Scholar 

  20. Mark W, Junru W, Jen-Fu, C . Ultrasound-induced cell lysis and sonoporation enhanced by contrast agents. J Acoust Soc Am. 1999;105:2951–2957.

    Article  Google Scholar 

  21. Evan CU, Evan H, Mani V, Terry OM, Thomas M . Local drug and gene delivery through microbubbles. Prog Cardiovasc Dis. 2001;44:45–54.

    Article  Google Scholar 

  22. Takahashi M, Sato T, Sagawa T, et al. E1B-55K-deleted adenovirus expressing E1A-13S by AFP-enhancer/promoter is capable of highly specific relation in AFP-producing hepatocellular carcinoma and eradication of established tumor. Mol Ther. 2002;5:627–634.

    Article  CAS  PubMed  Google Scholar 

  23. Alshad SL, Betty C, Jian MN, et al. Anti-tumor efficacy of human angiostatin using liver-mediated adeno-associated virus gene therapy. Mol Ther. 2004;9:56–66.

    Article  Google Scholar 

  24. Katsunori S, Oliver E, Chryssanthi K, Yun-Sheng T, Savio LCW . Oncolysis of multifocal hepatocellular carcinoma in the rat liver by hepatic artery infusion of vesicular stomatitis virus. Mol Ther. 2004;9:368–376.

    Article  Google Scholar 

  25. Tada H, Maron DJ, Choi EA, et al. Systemic IFN-β gene therapy results in long-term survival in mice with established colorectal liver metastases. J Clin Invest. 2001;108:83–95.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Volker S, Miguel B, Lin W, et al. Adenovirus-mediated CD40 ligand gene therapy in a rat model of orthotopic hepatocellular carcinoma. Hepatology. 2001;34:72–81.

    Article  Google Scholar 

  27. Ruian X, Xueying S, Lai-Yin T, et al. Long-term expression of angiostatin suppresses metastatic liver cancer in mice. Hepatology. 2003;37:1451–1460.

    Article  Google Scholar 

  28. Ben-Gary H, Robin LM, Todd R, Martin LL, Ronald GC . Systemic interleukin-6 responses following administration of adenovirus gene transfer vectors to humans by different routes. Mol Ther. 2002;6:287–297.

    Article  PubMed  Google Scholar 

  29. Sorin A, Ulrich ML, Irina S, et al. Adenoviral gene transfer of tumor necrosis factor-related apoptosis-inducing ligand overcomes an impaired response of hepatoma cells but causes severe apoptosis in primary human hepatocytes. Cancer Res. 2003;63:2369–2372.

    Google Scholar 

  30. Patricia Y, AR Davis, MJ Hicks, NS Templeton, Benner MK . Liposomal enhancement of the antitumor activity of conditionally replication-competent adenoviral plasmids. Mol Ther. 2004;9:489–495.

    Article  Google Scholar 

  31. Kim KS, Kim HS, Park JS, Kwon YG, Park YS . Inhibition of B16BL6 tumor progression by coadministration of recombinant angiostatin K1-3 and endostatin genes with cationic liposomes. Cancer Gene Ther. 2004;11:441–449.

    Article  PubMed  Google Scholar 

  32. Moret-Tatay I, Diaz J, Marco FM, Crespo A, Alino SF . Complete tumor prevention by engineered tumor cell vaccines employing nonviral vectors. Cancer Gene Ther. 2003;10:887–897.

    Article  CAS  PubMed  Google Scholar 

  33. van Zanten J, Doornbos-Van der Meer B, Audouy S, Kok RJ, de Leij L . A nonviral carrier for targeted gene delivery to tumor cells. Cancer Gene Ther. 2004;11:156–164.

    Article  CAS  PubMed  Google Scholar 

  34. Hajri A, Wack S, Lehn P, et al. Combined suicide gene therapy for pancreatic peritoneal carcinomatosis using BGTC liposomes. Cancer Gene Ther. 2004;11:16–27.

    Article  CAS  PubMed  Google Scholar 

  35. Sakurai F, Terada T, Maruyama M, et al. Therapeutic effect of intravenous delivery of lipoplexes containing the interferon-beta gene and poly I: poly C in a murine lung metastasis model. Cancer Gene Ther. 2003;10:661–668.

    Article  CAS  PubMed  Google Scholar 

  36. Matus S, Frank CM, Richard EC, Claudia Z, Isaiah JF, Michael A . Bone marrow-derived mesenchymal stem cells as vehicles for interferon-β delivery into tumors. Cancer Res. 2002;62:3603–3608.

    Google Scholar 

  37. Takaoka A, Hayakawa S, Yanai H, et al. Integration of interferon-α/β signalling to p53 responses in tumour suppression and antiviral defense. Nature. 2003;424:516–523.

    Article  CAS  PubMed  Google Scholar 

  38. Scott MF, Rajagopal R, Munish S, Anupama M, Andrea KJ, Aizen JM . The role of cytokines in mediating the bystander effect using HSV-TK xenogeneic cells. Cancer Lett. 1995;92:167–174.

    Article  Google Scholar 

  39. Hall SJ, Canfield SE, Yan Y, Hassen W, Selleck WA, Chen S-H . A novel bystander effect involving tumor cell-derived Fas and FasL interactions following Ad.HSV-tk and Ad.mIL-12 gene therapies in experimental prostate cancer. Gene Therapy. 2002;9:511–517.

    Article  CAS  PubMed  Google Scholar 

  40. Tachibana K, Tachibana S . Albumin microbubble echo-contrast material as an enhancer for ultrasound accelerated thrombolysis. Circulation. 1995;92:1148–1150.

    Article  CAS  PubMed  Google Scholar 

  41. Markus FW, Christiane T, Malgorzata K, et al. Specific systemic nonviral gene delivery to human hepatocellular carcinoma xenografts in SCID mice. Hepatology. 2002;36:1106–1114.

    Article  Google Scholar 

  42. Marcus F, John FB, Sandra P, Jesse ES, Gordhan LP, Stephen GZ . Transfection of mammalian cells with plasmid DNA by scrape loading and sonication loading. Proc Natl Acad Sci USA. 1987;84:8463–8467.

    Article  Google Scholar 

Download references

Acknowledgements

We thank Ms Anne Broillet (Bracco Research SA) for supplying the contrast agent (BR14) and Riken BRC for providing the pCMV(Hu beta) and pCAGGS-incorporated LacZ genes. We are also grateful to Ms Tomoko Murase (Department of Surgery II, Nagoya University School of Medicine) for her technical support. These studies were supported by a Grant-in-Aid for Scientific Research on Priority Areas (B)(2) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

Author information

Authors and Affiliations

  1. Department of Surgery II, Nagoya University School of Medicine, Nagoya, 466-8550, Japan

    Yoshikazu Sakakima & Akimasa Nakao

  2. Department of Surgery, National East Nagoya Hospital, Nagoya, 465-8620, Japan

    Shuji Hayashi

  3. Department of Surgery, National Toyohashi Hospital, Toyohashi, 441-8585, Japan

    Yoshikazu Yagi

  4. Department of Equipment Center for Research and Education, Nagoya University Graduate School of Medicine, Nagoya, 466-8550, Japan

    Akemi Hayakawa

  5. Department of Anatomy, School of Medicine, Fukuoka University, Fukuoka, 814-0180, Japan

    Katsuro Tachibana

Authors
  1. Yoshikazu Sakakima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuji Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoshikazu Yagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Akemi Hayakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Katsuro Tachibana
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Akimasa Nakao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoshikazu Sakakima.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sakakima, Y., Hayashi, S., Yagi, Y. et al. Gene therapy for hepatocellular carcinoma using sonoporation enhanced by contrast agents. Cancer Gene Ther 12, 884–889 (2005). https://doi.org/10.1038/sj.cgt.7700850

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.cgt.7700850

Keywords

This article is cited by

Search

Advanced search

Quick links