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Inhibition of hepatocellular carcinoma growth and angiogenesis by dual silencing of NET-1 and VEGF

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Abstract

Simultaneous silencing of multiple up-regulated genes is an attractive and viable strategy to treat many incurable diseases including cancer. Herein we used dual gene targeted siRNA (DGT siRNA) conjugate composed of NET-1 and VEGF siRNA sequences in the same backbone could inhibit growth and angiogenesis HCC. DGT siRNA showed a further down regulation on VEGF mRNA and protein levels compared with NET-1 siRNA or VEGF siRNA, but not on NET-1 expression. It also exhibited greater suppression on proliferation and trigger of apoptosis in HepG2 cells than NET-1 siRNA or VEGF siRNA; this could be explained by the significant down regulation of cyclin D1 and Bcl-2. A lower level of ANG2 mRNA and protein was detected in HUVEC cultured with supernatant of HepG2 cells treated with DGT siRNA than that of VEGF siRNA or NET-1 siRNA, resulting in much more inhibited angiogenesis of HUVEC. Tumor growth was inhibited and microvessel density dropped in the xenograft tumor models compared to the untreated controls. NET-1 and VEGF silencing play a key role in inhibiting hepatocellular cell proliferation, promoting apoptosis, and reducing angiogenesis. Simultaneous silencing of NET-1 and VEGF using DGT siRNA construct may provide an advantageous alternative in development of therapeutics for Hepatocellular carcinoma.

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Acknowledgments

This study was supported by fundings: from the Advanced Academic Program Development of Jiangsu Higher Education Institutions, Foundation of the Ministry of Health, Jangsu provience, China (No.H201052), the Science Foundation of Nantong City, Jiangsu province, China (No.K2009060), and Foundation of medical school, Nantong university, Jangsu province, China (No.YXY-200908), respectively. We thank Biomics Biotechnologies Co., Ltd. (Nantong, Jiangsu, China) for kindly providing synthesis of siRNAs and shRNAs.

Conflict of interest

All the authors declared no conflict of interest.

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Authors and Affiliations

  1. Department of Pathological Anatomy, Nantong University, Nantong, People’s Republic of China

    Yuan-Yuan Wu, Li Chen, Gui-Lan Wang, Jia-Ming Zhou & Jing Qin

  2. Comparative Medicine Institute, Nantong University, Nantong, People’s Republic of China

    Yuan-Yuan Wu

  3. Department of Pathology, Nantong Tumor Hospital, Nantong, People’s Republic of China

    Yi-Xin Zhang & Song He

  4. Biomics (Nantong) Co., Ltd, Nantong, People’s Republic of China

    Yuan-Yuan Zhu

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  1. Yuan-Yuan Wu
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  2. Li Chen
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Correspondence to Li Chen.

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Li Chen: Joint first author

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Wu, YY., Chen, L., Wang, GL. et al. Inhibition of hepatocellular carcinoma growth and angiogenesis by dual silencing of NET-1 and VEGF. J Mol Hist 44, 433–445 (2013). https://doi.org/10.1007/s10735-012-9480-5

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  • DOI: https://doi.org/10.1007/s10735-012-9480-5

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