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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 Therapy volume 15, pages 776–786 (2008)Cite this article

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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Authors and Affiliations

  1. Division of Basic Medicine I, Department of Anatomy and Cell Biology, Osaka Medical College, Takatsuki, Osaka, Japan

    M-A Shibata & Y Otsuki

  2. High-Tech Research Center, Osaka Medical College, Takatsuki, Osaka, Japan

    M-A Shibata, E Shibata & Y Otsuki

  3. Laboratory Animal Center, Osaka Medical College, Takatsuki, Osaka, Japan

    J Morimoto

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  1. M-A Shibata
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Correspondence to M-A Shibata or Y Otsuki.

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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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