title: Gene-silencing effects of anti-survivin siRNA delivered by RGDV-functionalized nanodiamond carrier in the breast carcinoma cell line MCF-7.
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 60-110 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.
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Yanzhao
authors.initials: Y
authors.lastname: Bi
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Chunying
authors.initials: C
authors.lastname: Cui
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Lulu
authors.initials: L
authors.lastname: Ren
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Xueyun
authors.initials: X
authors.lastname: Jiang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry*; D020868:Gene Silencing; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000172:deficiency* / Q000235:genetics*; D061986:MCF-7 Cells; D008297:Male; D051379:Mice; D058612:Nanodiamonds / Q000737:chemistry*; D050997:Nanomedicine; D009842:Oligopeptides / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics*; D000077022:Survivin
chemical_list: C107489:BIRC5 protein, human; D004337:Drug Carriers; D051034:Inhibitor of Apoptosis Proteins; D058612:Nanodiamonds; D009842:Oligopeptides; D034741:RNA, Small Interfering; D000077022:Survivin; C047981:arginyl-glycyl-aspartic acid
keywords: NDCONH(CH2)2NH-VDGR; RNAi; siRNA carrier; survivin