Cationic liposome-mediated transfection of CD40 ligand gene inhibits hepatic tumor growth of hepatocellular carcinoma in mice.

Abstract

OBJECTIVE:

To evaluate the efficacy of cationic liposome-mediated CD40 ligand (CD40L) gene therapy for hepatocellular carcinoma.

METHODS:

1x10(6) of parental H22 cells or H22 cells transfected with the expression vector containing murine CD40L cDNA encoding the entire coding region (pcDNA3.1(+)-mCD40L) were inoculated subcutaneously into the left flanks of syngenic BALB/C mice. The tumor-bearing mice (tumor nodules 10 mm in maximal diameter) received the treatment of the intratumoral injection of pcDNA3.1(+)-mCD40L/Transfectam, pcDNA3.1(+), or phosphate-buffered saline (PBS), or no treatment. The mice were monitored for tumor growth weekly. We examined mCD40L messenger ribonucleic acid (mRNA) expression by reverse transcription polymerase chain reaction (RT-PCR) and the histologic changes in tumors at two weeks after intratumoral injection using immunohistochemical staining of tumor tissues.

RESULTS:

All mice inoculated with parental H22 cells developed a tumor subcutaneously, and the tumor size increased progressively within three weeks. However, the mice receiving H22-CD40L cells exhibited complete regression of the tumor two weeks after tumor cell inoculation. The tumor-bearing animals with the treatment of pcDNA3.1(+) or PBS, or without treatment had progressive tumor growth, while those mice treated with pcDNA3.1(+)-mCD40L exhibited a significant inhibition of tumor growth. RT-PCR analysis showed that 783-bp fragments corresponding to the mCD40L mRNA were amplified only from pcDNA3.1(+)-mCD40L treated tumors. The tumor samples from pcDNA3.1(+)-mCD40L-treated mice showed significant lymphocyte infiltration, apoptotic bodies, and confluent necrosis in the tumor tissues.

CONCLUSION:

The tumorigenicity of CD40L-expressing cells was abrogated when the cells were implanted subcutaneously. In vivo gene therapy of established liver tumor nodules in mice by the intratumoral injection of pcDNA3.1(+)-mCD40L led to significant tumor inhibition. There was mCD40L mRNA expression in the tissues from pcDNA3.1(+)-mCD40L-treated tumors. The intratumoral injection of pcDNA3.1(+)-mCD40L induced a strong inflammatory, mainly lymphocytic infiltration of the tumor, and increased the necrotic rate of the neoplastic cells.

• Authors: Yong-fang Jiang, Jing Ma, Yan He, Yong-hong Zhang, Yun Xu, Guo-zhong Gong
• Journal: Journal of Zhejiang University. Science. B (J Zhejiang Univ Sci B) Vol. 10 Issue 1 Pg. 7-13 (Jan 2009) ISSN: 1673-1581 [Print] China
• PMID: 19198017 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Cations
• Liposomes
• CD40 Ligand

Topics

• Animals
• CD40 Ligand (genetics, therapeutic use)
• Carcinoma, Hepatocellular (pathology, therapy)
• Cations
• Cell Proliferation
• Female
• Genetic Therapy (methods)
• Liposomes (chemistry)
• Liver Neoplasms (pathology, therapy)
• Mice
• Mice, Inbred BALB C
• Transfection (methods)
• Treatment Outcome