Abstract
Vascular endothelial growth factor (VEGF) is up-regulated in the vast majority of human tumors. The up-regulation of VEGF not only plays important roles in tumor angiogenesis, but also provides a target for tumor treatment with small interfering RNA (siRNA) that targets VEGF; however, it is unclear whether a quite high up-regulation of VEGF will affect the efficiency of RNA interference strategies targeting VEGF. A high level expression of VEGF was found in CNE cells from a nasopharyngeal carcinoma cell line. In this study, we investigate whether VEGF-specific siRNAs can effectively suppress VEGF expression in CNE cells, and study the methods for the use of VEGF-specific siRNAs as potential therapeutic agents. CNE cells with high VEGF expression induced by hypoxia were transfected with VEGF-specific siRNAs. The expression of VEGF was effectively suppressed by VEGF-specific siRNAs, measured by ELISA, Western blot analysis and RT-PCR. Furthermore, experiments in nude mice bearing nasopharyngeal carcinoma xenograft were initiated 5 d after injection of CNE cells. VEGF-specific siRNAs were modified with 2′-deoxy, then injected into the tumors, and a liposome-mediated siRNA transfection system and ultrasound exposure were used to help delivery of the siRNAs. Tumor growth was reduced significantly after 3 weeks’ treatment. These studies suggest that VEGF-specific siRNAs still can effectively suppress VEGF expression even in tumor cell lines with a relatively high level of VEGF expression, such as CNE, and VEGF-specific siRNAs modified with 2′-deoxy can be used as potential agents for tumor therapy.
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References
Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med, 1995, 1: 27–31
Zetter B R. Angiogenesis and tumor metastasis. Annu Rev Med, 1998, 49: 407–424
Ferrara N, Davis-Smyth T. The biology of vascular endothelial growth factor. Endocr Rev, 1997, 18: 4–25
Ferrara N, Gerber H P, LeCouter J. The biology of VEGF and its receptors. Nat Med, 2003, 9: 669–676
Raab-Traub N, Flynn K. The structure of the termini of the Epstein-Barr virus as a marker of clonal cellular proliferation. Cell, 1986, 47: 883–889
Pathmanathan R, Prasad U, Sadler R, et al. Clonal proliferations of cells infected with Epstein-Barr virus in preinvasive lesions related to nasopharyngeal carcinoma. N Engl J Med, 1995, 333: 693–698
Murono S, Inoue H, Tanabe T, et al. Induction of cyclooxygenase-2 by Epstein-Barr virus latent membrane protein 1 is involved in vascular endothelial growth factor production in nasopharyngeal carcinoma cells. Proc Natl Acad Sci USA, 2001, 98: 6905–6910
Lieberman J, Song E, Lee S K, et al. Interfering with disease: opportunities and roadblocks to harnessing RNA interference. Trends Mol Med, 2003, 9: 397–403
Mello CC, Conte D Jr. Revealing the world of RNA interference. Nature, 2004, 431: 338–342
Elbashir S M, Harborth J, Lendeckel W, et al. Duplexes of 21-nucleotide RNAs mediate interference in cultured mammalian cells. Nature, 2001, 411: 494–498
Reynolds A, Leake D, Boese Q, et al. Rational siRNA design for RNA interference. Nat Biotechnol, 2004, 22: 326–330
Takei Y, Kadomatsu K, Yuzawa Y, et al. A small interfering RNA targeting vascular endothelial growth factor as cancer therapeutics. Cancer Res, 2004, 64: 3365–3370
Burfeind P, Chernicky C L, Rininsland F, et al. Antisense RNA to the type I insulin-like growth factor receptor suppresses tumor growth and prevents invasion by rat prostate cancer cells in vivo. Proc Natl Acad Sci USA, 1996, 93: 7263–7268
Hui E P, Chan A T, Pezzella F, et al. Coexpression of hypoxia-inducible factors 1alpha and 2alpha, carbonic anhydrase IX, and vascular endothelial growth factor in nasopharyngeal carcinoma and relationship to survival. Clin Cancer Res, 2002, 8: 2595–2604
Verma I M, Somia N. Gene therapy: promises, problems and prospects. Nature 1997, 389: 239–242
Niidome T, Huang L. Gene therapy progress and prospects: nonviral vectors. Gene Therapy, 2002, 9: 1647–1652
Li S, Huang L. Nonviral gene therapy: promises and challenges. Gene Therapy, 2000, 7: 31–34
Tachibana K, Uchida T, Ogawa K, et al. Induction of cell-membrane porosity by ultrasound. Lancet, 1999, 353: 1409–1411
Gambihler S, Delius M, Ellwart J W. Permeabilization of the plasma membrane of L1210 mouse leukemia cells using lithotripter shock waves. J Membr Biol, 1994, 141: 267–275
Lawrie A, Brisken A F, Francis S E, et al. Ultrasound enhances reporter gene expression after transfection of vascular cells in vitro. Circulation, 1999, 99: 2617–2620
Bekeredjian R, Chen S, Frenkel P A, et al. Ultrasound-targeted microbubble destruction can repeatedly direct highly specific plasmid expression to the heart. Circulation, 2003, 108: 1022–1026
Chiu Y L, Rana T M. SiRNA function in RNAi: A chemical modification analysis. RNA, 2003, 9: 1034–1048
Hall A H S, Wan J, Shaughnessy E E, et al. RNA interference using boranophosphate siRNAs: Structure-activity relationships. Nucleic Acids Res, 2004, 32: 5991–6000
Braasch D, Jensen S, Liu Y, et al. RNA interference in mammalian cells by chemically-modified RNA. Biochemistry, 2003, 42: 7967–7975
Layzer J M, Mccaffrey A P, Tanner A K, et al. In vivo activity of nucleaseresistant siRNAs. RNA, 2004, 10: 766–771
Amarzguioui M, Holen T, Babaie E, et al. Tolerance for mutations and chemical modifications in a siRNA. Nucleic Acids Res, 2003, 31: 589–595
Czauderna F, Fechtner M, Dames S. Structural variations and stabilizing modifications of synthetic siRNAs in mammalian cells. Nucleic Acids Res, 2003, 31: 2705–2716
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Supported by Natural Science Foundation of Guangdong Province, China (Grant No. 012402)
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Chen, S., Gao, G., Chen, W. et al. VEGF-specific siRNAs modified with 2′-deoxy effectively suppress VEGF expression and inhibit growth of nasopharyngeal carcinoma xenograft in a mouse model. Sci. China Ser. C-Life Sci. 51, 104–110 (2008). https://doi.org/10.1007/s11427-008-0020-1
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DOI: https://doi.org/10.1007/s11427-008-0020-1