A U87‑EGFRvIII cell‑specific aptamer mediates small interfering RNA delivery

Zhang,Xingmei; Liang,Huiyu; Tan,Yan; Wu,Xidong; Li,Shuji; Shi,Yusheng

doi:10.3892/br.2014.276

A U87‑EGFRvIII cell‑specific aptamer mediates small interfering RNA delivery

Authors:
- Xingmei Zhang
- Huiyu Liang
- Yan Tan
- Xidong Wu
- Shuji Li
- Yusheng Shi
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Affiliations: Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Radiation Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: May 15, 2014 https://doi.org/10.3892/br.2014.276
Pages: 495-499

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Abstract

U87‑EGFRvIII is a U87 glioma cell line that overexpresses epidermal growth factor receptor variant III (EGFRvIII). In the present study, we investigated whether a DNA aptamer selected against U87‑EGFRvIII using cell‑based systematic evolution of ligands by exponential enrichment (cell‑SELEX) could deliver c‑Met small interfering RNA (siRNA) into U87‑EGFRvIII cells and silence the targeted gene expression. The selected biotinylated aptamer (BA) was coupled to biotinylated c‑Met siRNA by streptavidin to deliver siRNA into U87‑EGFRvIII cells. c‑Met siRNA, transfected with lipofectamine 2000, served as a positive control, while control siRNA, transferred with BA, served as a negative control. Western blotting was performed to detect changes in the c‑Met protein expression, and MTT and Annexin V‑fluorescein isothiocyanate/propidium iodine assays were used to determine changes in the proliferation and apoptosis of U87‑EGFRvIII cells, respectively. Similar to the liposome‑mediated group, U87‑EGFRvIII cells that were transfected with BA‑c‑Met siRNA experienced a significant decrease in the c‑Met protein expression (P<0.05). There were also significant increases in the apoptotic rate (P<0.05) and inhibition rate of cell growth (P<0.01) compared with the negative control group, indicating that BA could deliver c‑Met siRNA into U87‑EGFRvIII and result in target gene silencing. In conclusion, the results demonstrated that this DNA aptamer, obtained through cell‑SELEX, can be used as an efficient and targeted carrier for siRNA delivery, providing a novel approach and strategy for the targeted combination therapy of glioblastoma.

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1	Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K and Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411:494–498. 2001. View Article : Google Scholar : PubMed/NCBI
2	Burnett JC and Rossi JJ: RNA-based therapeutics: current progress and future prospects. Chem Biol. 19:60–71. 2012. View Article : Google Scholar : PubMed/NCBI
3	Tuerk C and Gold L: Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science. 249:505–510. 1990. View Article : Google Scholar
4	Sefah K, Shangguan D, Xiong X, O’Donoghue MB and Tan W: Development of DNA aptamers using Cell-SELEX. Nature Protoc. 5:1169–1185. 2010. View Article : Google Scholar : PubMed/NCBI
5	Gan HK, Kaye AH and Luwor RB: The EGFRvIII variant in glioblastoma multiforme. J Clin Neurosci. 16:748–754. 2009. View Article : Google Scholar : PubMed/NCBI
6	Liu W, Fu Y, Xu S, et al: c-Met expression is associated with time to recurrence in patients with glioblastoma multiforme. J Clin Neurosci. 18:119–121. 2011. View Article : Google Scholar : PubMed/NCBI
7	Chu TC, Twu KY, Ellington AD and Levy M: Aptamer mediated siRNA delivery. Nucleic Acids Res. 34:e732006. View Article : Google Scholar : PubMed/NCBI
8	McNamara JO II, Andrechek ER, Wang Y, et al: Cell type-specific delivery of siRNAs with aptamer-siRNA chimeras. Nat Biotechnol. 24:1005–1015. 2006. View Article : Google Scholar : PubMed/NCBI
9	Dassie JP, Liu XY, Thomas GS, et al: Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat Biotechnol. 27:839–849. 2009. View Article : Google Scholar : PubMed/NCBI
10	Zhou J, Li H, Zhang J, Piotr S and Rossi J: Development of cell-type specific anti-HIV gp120 aptamers for siRNA delivery. J Vis Exp. 52:29542011.PubMed/NCBI
11	Wheeler LA, Trifonova R, Vrbanac V, et al: Inhibition of HIV transmission in human cervicovaginal explants and humanized mice using CD4 aptamer-siRNA chimeras. J Clin Invest. 121:2401–2412. 2011. View Article : Google Scholar : PubMed/NCBI
12	Zhou J, Swiderski P, Li H, et al: Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells. Nucleic Acids Res. 37:3094–3109. 2009. View Article : Google Scholar : PubMed/NCBI
13	Thiel KW, Hernandez LI, Dassie JP, et al: Delivery of chemo-sensitizing siRNAs to HER2+-breast cancer cells using RNA aptamers. Nucleic Acids Res. 40:6319–6337. 2012.PubMed/NCBI
14	Tan Y, Shi YS, Wu XD, et al: DNA aptamers that target human glioblastoma multiforme cells overexpressing epidermal growth factor receptor variant III in vitro. Acta Pharmacol Sin. 34:1491–1498. 2013. View Article : Google Scholar
15	Zhan Y and O’Rourke DM: SHP-2-dependent mitogen-activated protein kinase activation regulates EGFRvIII but not wild-type epidermal growth factor receptor phosphorylation and glioblastoma cell survival. Cancer Res. 64:8292–8298. 2004. View Article : Google Scholar
16	Nicholas MK, Lukas RV, Chmura S, Yamini B, Lesniak M and Pytel P: Molecular heterogeneity in glioblastoma: therapeutic opportunities and challenges. Semin Oncol. 38:243–253. 2011. View Article : Google Scholar : PubMed/NCBI
17	Shangguan D, Cao ZC, Li Y and Tan W: Aptamers evolved from cultured cancer cells reveal molecular differences of cancer cells in patient samples. Clin Chem. 53:1153–1155. 2007. View Article : Google Scholar : PubMed/NCBI
18	Huang YF, Shangguan D, Liu H, et al: Molecular assembly of an aptamer-drug conjugate for targeted drug delivery to tumor cells. Chembiochem. 10:862–868. 2009. View Article : Google Scholar : PubMed/NCBI
19	Wu Y, Sefah K, Liu H, Wang R and Tan W: DNA aptamer-micelle as an efficient detection/delivery vehicle toward cancer cells. Proc Natl Acad Sci USA. 107:5–10. 2010. View Article : Google Scholar : PubMed/NCBI
20	Zhang K, Sefah K, Tang L, et al: A novel aptamer developed for breast cancer cell internalization. ChemMedChem. 7:79–84. 2012. View Article : Google Scholar : PubMed/NCBI
21	Thiel KW and Giangrande PH: Intracellular delivery of RNA-based therapeutics using aptamers. Ther Deliv. 1:849–861. 2010. View Article : Google Scholar : PubMed/NCBI
22	McCarty JH: Glioblastoma resistance to anti-VEGF therapy: has the challenge been MET? Clin Cancer Res. 19:1631–1633. 2013. View Article : Google Scholar : PubMed/NCBI
23	Peruzzi B and Bottaro DP: Targeting the c-Met signaling pathway in cancer. Clin Cancer Res. 12:3657–3660. 2006. View Article : Google Scholar : PubMed/NCBI
24	Engelman JA, Zejnullahu K, Mitsudomi T, et al: MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science. 316:1039–1043. 2007. View Article : Google Scholar : PubMed/NCBI
25	Pillay V, Allaf L, Wilding AL, et al: The plasticity of oncogene addiction: implications for targeted therapies directed to receptor tyrosine kinases. Neoplasia. 11:448–458, 2 p following 458. 2009.PubMed/NCBI
26	Huang PH, Mukasa A, Bonavia R, et al: Quantitative analysis of EGFRvIII cellular signaling networks reveals a combinatorial therapeutic strategy for glioblastoma. Proc Natl Acad Sci USA. 104:12867–12872. 2007. View Article : Google Scholar : PubMed/NCBI