Gene-silencing effects of anti-survivin siRNA delivered by RGDV-functionalized nanodiamond carrier in the breast carcinoma cell line MCF-7.
Gene-silencing effects of anti-survivin siRNA delivered by RGDV-functionalized nanodiamond carrier in the breast carcinoma cell line MCF-7.
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RGDV 功能化纳米金刚石载体递送的抗生存素 siRNA 在乳腺癌细胞系 MCF-7 中的基因沉默作用
DOI：
10.2147/ijn.s117611
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发表时间：
2016
影响因子：
8
通讯作者：
Jiang X
中科院分区：
文献类型：
--
作者：
Bi Y;Zhang Y;Cui C;Ren L;Jiang X
纳米金刚石（ND）是一种具有独特物理、化学和生物学特性的非病毒小干扰RNA（siRNA）载体材料。在我们前期的工作中，已经证明ND可以有效地将siRNA导入细胞，并下调所需蛋白的表达。然而，利用ND合成高效的肿瘤靶向载体仍然是一个挑战。本研究合成了一种新型的siRNA载体NDCONH（CH 2）2NH-VDGR，并采用傅里叶变换红外光谱、透射电镜、扫描电镜、凝胶阻滞法、差示扫描量热法、共聚焦显微镜、释放试验、实时荧光定量PCR（PCR）、酶联免疫吸附试验（ELISA）、流式细胞术、细胞毒性测定和体外和体内基因沉默功效测定。采用Annexin V-异硫氰酸荧光素/碘化丙啶染色法，通过流式细胞仪检测NDCONH（CH 2）2NH-VDGR/survivin-siRNA诱导肿瘤细胞凋亡的机制。制备了粒径为60-110 nm、zeta电位为35.65±3.90 mV的NDCONH（CH 2）2NH-VDGR/survivin-siRNA纳米粒。实时荧光定量PCR检测结果显示，Survivin mRNA的表达量下降至46.77%± 6.3%。ELISA检测Survivin蛋白表达下调至48.49%± 2.25%。MTT法显示NDCONH（CH 2）2NH-VDGR/survivin-siRNA对MCF-7细胞增殖有抑制作用。由此可见，survivin siRNA能够稳定地传递、转运和释放，有利于提高基因沉默效果。因此，NDCONH（CH 2）2NH-VDGR作为siRNA载体，有望应用于siRNA载体系统和肿瘤治疗中。
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 60–110 nm diameter and 35.65±3.90 mV 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.
影响因子：
14.9
作者：
Abe T;Goda K;Futami K;Furuichi Y
通讯作者：
Furuichi Y