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RNA interference (RNAi)-mediated vascular endothelial growth factor-C (VEGF-C) reduction interferes with lymphangiogenesis and enhances Epirubicin sensitivity of breast cancer cells

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Abstract

It has been reported that over-expression of vascular endothelial growth factor-C (VEGF-C) in tumors leads to increased lymphangiogenesis and resistance to chemotherapy. Therefore, we hypothesized that VEGF-C would be a good molecular target for cancer gene therapy. In this study, we silenced the expression of VEGF-C with the highly specific post-transcriptional suppression of RNA interference (RNAi) in human breast cancer MCF-7 cell line. The expression of VEGF-C was examined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), and the effect of plasmid on human lymphatic endothelial cells (HLECs) in vitro was analyzed by migration and 3-(4, 5-dimethylt-hiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The sensitivity to anticancer agents was evaluated by MTT and apoptosis assay, and apoptosis-related genes bcl-2/bax ratio was determined by Western Blotting. Results showed that of three siRNA-expressing vectors, P-1/siRNA most significantly suppressed the expression of VEGF-C mRNA and protein (38.1% of control and 117.8 ± 24.2 pg/ml, respectively) and interfered with proliferation and migration of HLECs in vitro. Moreover, transfection of VEGF-C/siRNA combined with Epirubicin markedly decreased breast cancer cells viability, reaching up to 38.5%, and increased apoptosis rate from 13.1% to 38.9%, as determined by decrease of bcl-2/bax ratio. In summary, VEGF-C would be a good molecular target, and a combination of Epirubicin and RNAi targeting VEGF-C could be an effective means for suppressing lymphatic metastasis and enhancing chemosensitivity of human breast cancer cells.

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Acknowledgment

We are very grateful to Prof. Zhao-Xi Ding of Shandong University in China for providing human lymphatic endothelial cells.

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Authors and Affiliations

  1. Department of Anatomy, School of Medicine, Shandong University, No. 44, WenHua Xi Road, JiNan, Shandong, 250012, P.R. China

    Ping Sun, Jie Gao, Yan-Li Liu, Lu-Wan Wei, Li-Ping Wu & Zhi-Yu Liu

  2. Department of Anatomy, MuDanJiang Medical College, No. 3, TongXiang Street, MuDanJiang, 157011, P.R. China

    Ping Sun

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  1. Ping Sun
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  2. Jie Gao
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  3. Yan-Li Liu
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Correspondence to Zhi-Yu Liu.

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Sun, P., Gao, J., Liu, YL. et al. RNA interference (RNAi)-mediated vascular endothelial growth factor-C (VEGF-C) reduction interferes with lymphangiogenesis and enhances Epirubicin sensitivity of breast cancer cells. Mol Cell Biochem 308, 161–168 (2008). https://doi.org/10.1007/s11010-007-9624-1

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  • DOI: https://doi.org/10.1007/s11010-007-9624-1

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