选择性识别平行性G-四联体的方酸类近红外荧光探针的研究
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摘要
G-四联体是抗肿瘤药物筛选的一个重要靶点.开发针对某些拓扑结构的G-四联体荧光探针对于研究其结构和生物学功能具有十分重要的意义.设计、合成了四个方酸类花菁荧光染料即CSTS,CSBE,CSEM和CSBM,并检测了其对不同类型DNA的选择性识别作用.结果表明,所合成的四个化合物在缓冲溶液中几乎没有荧光发射,加入正平行G-四联体之后荧光增强大约1 000倍;而加入反平行G-四链体或单双链DNA荧光仅仅增强几十倍,说明其可以特异性识别平行G-四联体.但流式实验结果显示,CSTS不能透过细胞膜,同时存在高荧光背景的缺点,因此无法应用于活体分析.而另外三个不带阴离子侧链的衍生物则容易进入细胞,进入细胞的难易顺序为CSBE>CSEM>CSBM>CSTS.高选择性、低背景荧光和易进入细胞等优点使CSBE具有作为近红外荧光探针检测生物样品中正平行G-四联体的潜力.
G-quadruplex is an important target of anti-tumor grug. The development of G-quadruplex fluorescent probes for certain topological structures is of great significance for studying their structure and biological functions. In this paper, four squaraine cyanine fluorescent dyes, namely CSTS, CSBE, CSEM and CSBM, were designed and synthesized. Their structures were determinated by ~1H NMR and MS. Little fluorescence emission was found in the buffer solution containing one of four squaraine cyanine fluorescent dyes. However, the fluorescence emission was enhanced approximately 1 000-fold after adding a normal parallel G-quadruplex and only enhanced several tens of times with the anti-parallel G-quadruplex fluorescence or single-stranded DNA. The results indicated that squaraine cyanine fluorescent dyes have the ability to selectively recognize of parallel G-quadruplex. Flow cytometry experiments showed that the CSTS could not invade the cells and had the disadvantages of high fluorescence background which could not be applied to mlecular pobes for bological imaging. The other three derivatives without an anionic side chain are easily accessible to cells. The order of difficulty in entering cells is following as: CSBE>CSEM>CSBM>CSTS. Due to the high selectivity, low fluorescence background and easy-to-access cells, CSBE have the potential as a near-infrared fluorescent probe to detect the positive parallel G-quadruplex in biological samples.
G-quadruplex is an important target of anti-tumor grug. The development of G-quadruplex fluorescent probes for certain topological structures is of great significance for studying their structure and biological functions. In this paper, four squaraine cyanine fluorescent dyes, namely CSTS, CSBE, CSEM and CSBM, were designed and synthesized. Their structures were determinated by ~1H NMR and MS. Little fluorescence emission was found in the buffer solution containing one of four squaraine cyanine fluorescent dyes. However, the fluorescence emission was enhanced approximately 1 000-fold after adding a normal parallel G-quadruplex and only enhanced several tens of times with the anti-parallel G-quadruplex fluorescence or single-stranded DNA. The results indicated that squaraine cyanine fluorescent dyes have the ability to selectively recognize of parallel G-quadruplex. Flow cytometry experiments showed that the CSTS could not invade the cells and had the disadvantages of high fluorescence background which could not be applied to mlecular pobes for bological imaging. The other three derivatives without an anionic side chain are easily accessible to cells. The order of difficulty in entering cells is following as: CSBE>CSEM>CSBM>CSTS. Due to the high selectivity, low fluorescence background and easy-to-access cells, CSBE have the potential as a near-infrared fluorescent probe to detect the positive parallel G-quadruplex in biological samples.
引文
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[4] BROOKS T A,KENDRICK S,HURLEY L.Making sense of G-quadruplex and i-motif functions in oncogene promoters [J].Febs Journal,2010,277(17):3459-3469.
[5] OU T M,LU Y J,TAN J H,et al.G-quadruplexes:Targets in anticancer drug design [J].Chemmedchem,2008,3(5):690-713.
[6] GEORGIADES S N,ABD KARIM N H,SUNTHARALINGAM K,et al.Interaction of metal complexes with G-quadruplex DNA [J].Angewandte Chemie-International Edition,2010,49(24):4020-4034.
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[8] DENG H,SUN M,WEI X P,et al.Electrochemical lead ion aptasensor based on G-quadruplex and gate-controlled signal amplification [J].Chinese Journal of Analytical Chemistry,2016,44(6):888-892.
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[12] VUMMIDI B R,ALZEER J,LUEDTKE N W.Fluorescent probes for G-quadruplex structures [J].Chembiochem,2013,14(5):540-558.
[13] ZUFFO M,DORIA F,BOTTI S,et al.G-quadruplex fluorescence sensing by core-extended naphthalene diimides [J].Biochimica Et Biophysica Acta-General Subjects,2017,1861(5):1303-1311.
[14] MIN H,ZHANG Z X,SHEN G L,et al.Near-infrared fluoresent probe of Azure a for determination of deoxyribonucleic acid [J].Chinese Journal of Analytical Chemistry,2007,35(6):890-892.
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[17] SONG B,ZHANG Q,MA W H,et al.The synthesis and photostability of novel squarylium indocyanine dyes [J].Dyes and Pigments,2009,82(3):396-400.
[18] WANG B S,FAN J L,SUN S G,et al.1-(Carbamoylmethyl)-3H-indolium squaraine dyes:synthesis,spectra,photo-stability and association with BSA [J].Dyes and Pigments,2010,85(1/2):43-50.
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[37] ZIELONKA J,JOSEPH J,SIKORA A,et al.Mitochondria-targeted triphenylphosphonium-based compounds:syntheses,mechanisms of action,and therapeutic and diagnostic applications [J].Chemical Reviews,2017,117(15):10043-10120.
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