Gene-silencing effects of anti-survivin siRNA delivered by RGDV-functionalized nanodiamond carrier in the breast carcinoma cell line MCF-7
| Autor: | Yanzhao Bi, Chunying Cui, Lulu Ren, Xueyun Jiang, Yifan Zhang |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine Small interfering RNA Materials science Survivin Biophysics Down-Regulation Pharmaceutical Science Apoptosis Bioengineering 02 engineering and technology Inhibitor of Apoptosis Proteins Nanodiamonds Flow cytometry Biomaterials Mice 03 medical and health sciences chemistry.chemical_compound International Journal of Nanomedicine In vivo Drug Discovery medicine Animals Humans MTT assay Electrophoretic mobility shift assay Gene Silencing Propidium iodide RNA Small Interfering Cytotoxicity Original Research Cell Proliferation Drug Carriers medicine.diagnostic_test Organic Chemistry General Medicine 021001 nanoscience & nanotechnology NDCONH(CH2)2NH-VDGR Molecular biology Nanomedicine 030104 developmental biology chemistry RNAi siRNA carrier MCF-7 Cells 0210 nano-technology Oligopeptides |
| Zdroj: | International Journal of Nanomedicine |
| ISSN: | 1178-2013 |
| DOI: | 10.2147/ijn.s117611 |
| Popis: | Yanzhao Bi, Yifan Zhang, Chunying Cui, Lulu Ren, Xueyun Jiang School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People’s Republic of China Abstract: Nanodiamond (ND) is a renowned material in nonviral small interfering RNA (siRNA) carrier field due to its unique physical, chemical, and biological properties. In our previous work, it was proven that ND could deliver siRNA into cells efficiently and downregulate the expression of desired protein. However, synthesizing a high-efficient tumor-targeting carrier using ND is still a challenge. In this study, a novel carrier, NDCONH(CH2)2NH-VDGR, was synthesized for siRNA delivery, and its properties were characterized with methods including Fourier transform infrared spectrometry, transmission electron microscopy, scanning electron microscopy, gel retardation assay, differential scanning calorimetry, confocal microscopy, releasing test, real-time polymerase chain reaction (PCR) assay, enzyme-linked immunosorbent assay (ELISA), flow cytometry, cytotoxicity assay, and gene-silencing efficacy assay in vitro and in vivo. The mechanism of NDCONH(CH2)2NH-VDGR/survivin-siRNA-induced tumor apoptosis was evaluated via flow cytometer assay using Annexin V–fluorescein isothiocyanate/propidium iodide staining method. The NDCONH(CH2)2NH-VDGR/survivin-siRNA nanoparticle with 60–110nm diameter and 35.65±3.90mV zeta potential was prepared. For real-time PCR assay, the results showed that the expression of survivin mRNA was reduced to 46.77%±6.3%. The expression of survivin protein was downregulated to 48.49%±2.25%, as evaluated by ELISA assay. MTT assay showed that NDCONH(CH2)2NH-VDGR/survivin-siRNA had an inhibitory effect on MCF-7 cell proliferation. According to these results, the survivin-siRNA could be delivered, transported, and released stably, which benefits in increasing the gene-silencing effect. Therefore, as an siRNA carrier, NDCONH(CH2)2NH-VDGR was suggested to be used in siRNA delivery system and in cancer treatments. Keywords: NDCONH(CH2)2NH-VDGR, siRNA carrier, RNAi, survivin |
| Databáze: | OpenAIRE |
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