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Antiangiogenic gene therapy for hepatocellular carcinoma using angiostatin gene
Hiroki Ishikawa1, Kazuhiko Nakao, Kojiro Matsumoto
1First Department of Internal Medicine, Nagasaki University School of Medicine, and Health Research Center, Nagasaki University, Nagasaki, Japan.
Abstract:
Recent studies have reported that antiangiogenic gene delivery into cancer cells inhibits growth of certain tumors in vivo. Hepatocellular carcinoma (HCC) is a hypervascular cancer, and antiangiogenic gene therapy might be suitable for HCC. In the present study, we investigated the antiangiogenic effects of angiostatin gene transduction into HCC both in vitro and in vivo. Angiostatin gene was cloned into a pSecTag2B mammalian expression vector to construct pSecTag2B-ANG. pSecTag2B or pSecTag2B-ANG were transfected into an HCC cell line, PLC/PRF/5, and then stable transfectants were obtained by Zeocin selection. pSecTag2B or pSecTag2B-ANG transfection did not alter the expression of vascular endothelial growth factor (VEGF), a potent angiogenic stimulator, or pigment epithelium-derived factor (PEDF), an angiogenic inhibitor, in PLC/PRF/5 cells. However, conditioned media (CM) derived from pSecTag2B-ANG-transfected PLC/PRF/5 cells (CM-ANG) suppressed the proliferation and migration of human umbilical vein endothelial cells (HUVEC) by 35% and 50%, respectively, relative to their effects on nontransfected cells. In in vivo experiments, pSecTag2B-ANG stable transfected (CM-Mock) and nontransfected cells (CM-N) were mixed at various proportions and the mixed cells were subcutaneously implanted into athymic mice. Suppression of tumor growth was noted in mice implanted with angiostatin gene-transfected cells, and such suppression was proportional with the percentage of transfected cells. Analysis of the vascular density in these tumors showed that the tumor growth suppression effect of angiostatin gene correlated with suppression of tumor vascularity. In conclusion, antiangiogenic gene therapy using angiostatin gene is potentially suitable for the treatment of patients with HCC.
Insights
Antiangiogenic gene therapy using angiostatin effectively inhibited hepatocellular carcinoma (HCC) growth in preclinical models. This approach suppressed tumor vascularity and growth, showing promise for HCC treatment.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) is a hypervascular malignancy.
- Antiangiogenic gene therapy shows potential for inhibiting tumor growth.
- Angiostatin is a known antiangiogenic factor.
Purpose of the Study:
- To investigate the antiangiogenic effects of angiostatin gene transduction in HCC.
- To evaluate the efficacy of angiostatin gene therapy in vitro and in vivo.
- To determine the impact on tumor vascularity and growth.
Main Methods:
- Cloning the angiostatin gene into a mammalian expression vector (pSecTag2B-ANG).
- Transfecting HCC cells (PLC/PRF/5) and establishing stable cell lines.
- Assessing effects on human umbilical vein endothelial cells (HUVEC) proliferation and migration.
- Implanting transfected HCC cells into athymic mice and analyzing tumor vascular density.
Main Results:
- Conditioned media from angiostatin-transfected HCC cells suppressed HUVEC proliferation and migration.
- Subcutaneous tumors in mice showed suppressed growth when implanted with angiostatin gene-transfected cells.
- Tumor growth suppression correlated with reduced tumor vascularity.
- VEGF and PEDF expression levels in HCC cells remained unaltered.
Conclusions:
- Angiostatin gene delivery demonstrates significant antiangiogenic effects against HCC.
- The study supports the potential of angiostatin gene therapy for HCC treatment.
- Reduced tumor vascularity is a key mechanism for growth inhibition.