Abstract
Cancer metastasis contributes significantly to cancer mortality and is facilitated by lymphangiogenesis and angiogenesis. Vascular endothelial growth factor-C (VEGF-C) and VEGF-A are involved in lymphangiogenesis and angiogenesis. To inhibit metastasis, combination therapy with vector-based small interfering RNA (siRNA) against VEGF-C and/or VEGF-A was conducted on murine metastatic mammary cancer. Syngeneic, inoculated, metastatic mammary cancers received direct intratumoral injection of plasmid siRNA vector targeting VEGF-C (psiRNA-VEGF-C), VEGF-A (psiRNA-VEGF-A), both VEGF-C and VEGF-A (both psiRNA-VEGF-C and psiRNA-VEGF-A vectors injected, referred to as the psiRNA-VEGF-C+A group) or a scrambled sequence (psiRNA-SCR) as control, once a week for 8 weeks. Gene electrotransfer was performed on the tumors after each injection. Tumor volume was significantly lower in the psiRNA-VEGF-A and the psiRNA-VEGF-C+A groups throughout the study. Lymph node metastasis was significantly less frequent in all therapeutic groups, whereas the multiplicity of lung metastases was significantly lower in the psiRNA-VEGF-C+A group only. All siRNA therapeutic groups showed a significant reduction in the number of dilated lymphatic vessels containing intraluminal cancer cells and microvessel density. Our data suggest that specific silencing of the VEGF-C or VEGF-A gene alone can inhibit lymph node metastasis. However, combination siRNA therapy targeting both VEGF-C and VEGF-A inhibits both lymph node and lung metastasis, rendering this combined therapy more beneficial than either alone. The observed anti-metastatic activity of siRNA-expressing vectors targeting VEGF-C or VEGF-A may be of high clinical significance in the treatment of metastatic breast cancer.
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Abbreviations
- RT-PCR:
-
reverse transcription-PCR
- shRNA:
-
short hairpin RNAs
- siRNA:
-
short interfering RNA
- VEGF:
-
vascular endothelial growth factor
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Acknowledgements
This investigation was supported by a Grant-in-Aid for Scientific Research (C)(2) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (no. 17591360 to MA Shibata) and, in part, by a High-Tech Research Center Grant to Osaka Medical College from MEXT. We also thank Ms Hidemi Hiyama and Mika Yoshida for their excellent secretarial assistance.
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Shibata, MA., Morimoto, J., Shibata, E. et al. Combination therapy with short interfering RNA vectors against VEGF-C and VEGF-A suppresses lymph node and lung metastasis in a mouse immunocompetent mammary cancer model. Cancer Gene Ther 15, 776–786 (2008). https://doi.org/10.1038/cgt.2008.43
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DOI: https://doi.org/10.1038/cgt.2008.43
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