Anti-angiogenic gene therapy for hepatocellular carcinoma mediated by microbubble-enhanced ultrasound exposure: an in vivo experimental study.

Abstract

OBJECTIVE:

The aim of this study was to assess the effect of anti-angiogenic gene therapy for hepatocelluar carcinoma (HCC) treated by microbubble-enhanced ultrasound exposure.

METHODS:

Forty C57BL/6J female mice were inoculated s.c. with Hepa1-6 tumor cell line. Herpes simplex virus thymidine kinase under the control of kinase domain-containing receptor (KDR, angiogenic growth factor's corresponding receptor) promoter was used. Plasmid DNA with or without microbubble contrast agent of SonoVuetrade mark was i.v. injected. Ultrasound (1 MHz, 2 W/cm(2), 5 min) was delivered to hepatic carcinomas in mice. The KDR-tk gene transfer was followed by ganciclovir (GCV) injection for 10 days and then the diameters of tumors were measured every 4 days till 28 days. The survivals of tumor-bearing mice were observed. PCR analysis and immunohistochemistry measurements revealed expression of the transfected gene. Transferase-mediated dUTP nick end labeling staining was used to detect apoptotic cells.

RESULTS:

Compared with the group treated by ultrasound alone, KDR-tk gene treatment treated by ultrasound combined with SonoVue restrained tumor growth and increased survival time of tumor-bearing mice; microvessel density in group mediated by ultrasound and SonoVue was significantly lower than that in group ultrasound alone (12.3 +/- 1.4 vs. 27.4 +/- 3.2, P < 0.05). An apoptosis index increased in the group treated by ultrasound and SonoVue compared with the group treated by ultrasound alone (25 +/- 3.6 vs. 36 +/- 3.8, P < 0.05), whereas there was no significant difference between group mediated by SonoVue alone and group phosphate-buffered saline alone (17 +/- 1.8 vs. 14 +/- 1.2, P>0.05).

CONCLUSIONS:

Gene therapy mediated by ultrasound exposure enhanced by a microbubble contrast agent may become a new treatment option for persistent HCC.

• Authors: Fang Nie, Hui-Xiong Xu, Ming-De Lu, Ying Wang, Qing Tang
• Journal: Journal of drug targeting (J Drug Target) Vol. 16 Issue 5 Pg. 389-95 (Jun 2008) ISSN: 1029-2330 [Electronic] England
• PMID: 18569283 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Contrast Media
• Phospholipids
• contrast agent BR1
• Thymidine Kinase
• Sulfur Hexafluoride

Topics

• Animals
• Carcinoma, Hepatocellular (blood supply, therapy)
• Cell Line, Tumor
• Contrast Media
• Female
• Genetic Therapy
• Immunohistochemistry
• Liver Neoplasms (blood supply, therapy)
• Mice
• Mice, Inbred C57BL
• Microbubbles
• Neoplasm Transplantation
• Neovascularization, Pathologic (therapy)
• Phospholipids
• Plasmids
• Reverse Transcriptase Polymerase Chain Reaction
• Simplexvirus (enzymology)
• Sulfur Hexafluoride
• Thymidine Kinase (genetics)
• Transfection
• Ultrasonics