雷鸣琳,孟雪莹,卢宏飞,孟雪明,赵艺卓,邢小静

• 1:南阳师范学院化学与制药工程学院

摘要(Abstract)：

人脱嘌呤/嘧啶核酸内切酶1(APE1)作为碱基切除修复(BER)途径中的关键酶，在肿瘤细胞中异常表达，是癌症诊断的生物标志物之一，因此开发高灵敏度的APE1活性检测方法具有重要的研究价值。基于双足DNA Walker较高的行走效率和稳定性，以核酸外切酶Ⅲ(ExoⅢ)为驱动力，通过其循环剪切金纳米颗粒(AuNPs)上连接的轨道链与双足DNA Walker形成的双链，释放荧光基团(FAM),实现信号放大，并利用AuNPs的荧光猝灭机制，开发了一种基于双足DNA Walker的荧光生物传感器，用于高灵敏度检测APE1。在最优检测条件下，APE1的活性在0.01～0.5 U/mL范围内与体系的荧光强度呈现良好线性关系，线性方程为F=173.00C_(APE1)+99.45,检测限为0.006 U/mL。本方法可用于血清模拟样品中APE1活性的检测，加标回收率为96.4%～98.5%。

关键词(KeyWords)： APE1;金纳米颗粒;双足DNA Walker;信号放大;生物传感器

Abstract:

Keywords:

基金项目(Foundation): 河南省科技攻关项目(252102320248);; 河南省青年骨干教师培育基金(2021GGJS123);; 河南省高等学校重点科研项目(24A150027)

作者(Author): 雷鸣琳,孟雪莹,卢宏飞,孟雪明,赵艺卓,邢小静

参考文献(References)：

• [1] CHOI Y D,JUNG J Y,BAEK M,et al.APE1 promotes pancreatic cancer proliferation through GFRα1/Src/ERK axis-cascade signaling in response to GDNF[J].International Journal of Molecular Sciences,2020,21(10):3586.
• [2] KLADOVA O A,ALEKSEEVA I V,SAPARBAEV M,et al.Modulation of the apurinic/ apyrimidinic endonuclease activity of human APE1 and of its natural polymorphic variants by base excision repair proteins[J].International Journal of Molecular Sciences,2020,21(19):7147.
• [3] FANTINI D,VASCOTTO C,DEGANUTO M,et al.APE1/Ref-1 regulates PTEN expression mediated by Egr-1[J].Free Radical Research,2008,42(1):20-29.
• [4] LAEV S S,SALAKHUTDINOV N F,LAVRIK O I.Inhibitors of nuclease and redox activity of apurinic/apyrimidinic endonuclease 1/redox effector factor 1 (APE1/Ref-1)[J].Bioorganic Medicinal Chemistry,2017,25(9):2531-2544.
• [5] ZHANG S H,HE L,DAI N,et al.Serum APE1 as a predictive marker for platinum-based chemotherapy of non-small cell lung cancer patients[J].Oncotarget,2016,7(47):77482-77494.
• [6] DAI N,CAO X J,LI M X,et al.Serum APE1 autoantibodies:a novel potential tumor marker and predictor of chemotherapeutic efficacy in non-small cell lung cancer[J].Plos One,2013,8(3):e58001.
• [7] KREKLAU E L,LIMP-FOSTER M,LIU N,et al.A novel fluorometric oligonucleotide assay to measure O 6-methylguanine DNA methyltransferase,methylpurine DNA glycosylase,8-oxoguanine DNA glycosylase and abasic endonuclease activities:DNA repair status in human breast carcinoma cells overexpressing methylpurine DNA glycosylase[J].Nucleic Acids Research,2001,29(12):2558-2566.
• [8] HAN J,ZHUO Y,CHAI Y Q,et al.Ultrasensitive electrochemical strategy for trace detection of APE-1 via triple signal amplification strategy[J].Biosensors and Bioelectronics,2013,41:116-122.
• [9] HOU T L,ZHU L,ZHANG X L,et al.Multiregion linear DNA Walker-mediated ultrasensitive electrochemical biosensor for miRNA detection[J] Analytical Chemistry,2022,94(29):10524-10530.
• [10] 张立燕，刘晓亭，杨亚允，等.DNA步行器在生物传感和活细胞成像领域的应用[J].中国科学：化学，2021,51(6):651-664.
• [11] YUAN A J,XIAO H,YANG F,et al.DNA Walker for signal amplification in living cells[J].Trends in Analytical Chemistry,2023,158:116870.
• [12] 黄惠，王江，霍冰洋，等.核酸介导信号放大光学生物传感器在食品污染物检测中的应用[J].分析试验室，2022,41(1):103-112.
• [13] 陈晶晶，郑斌，王瑜，等.DNA步行者在检测传感中的应用[J].分析试验室，2023,42(2):254-265.
• [14] WANG N,JIANG Y J,ZHANG X,et al.Nanosurface energy transfer indicating Exo Ⅲ-propelled stochastic 3D DNA Walkers for HIV DNA detection[J].Analyst,2021,146(5):1675-1681.
• [15] MA H M,CHEN L,LV J N,et al.The rate-limiting procedure of 3D DNA Walkers and their applications in tandem technology[J].Chemical Communications,2023,59(69):10330-10342.
• [16] HU H,ZHOU F,WANG B J,et al.Autonomous operation of 3D DNA Walkers in living cells for microRNA imaging[J].Nanoscale,2021,13(3):1863-1868.
• [17] LIU J W,LU Y.Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes[J].Nature Protocols,2006,1(1):246-252.
• [18] ZHAI J,LIU Y B,HUANG S,et al.A specific DNA-nanoprobe for tracking the activities of human apurinic/apyrimidinic endonuclease 1 in living cells[J].Nucleic Acids Research,2017,45(6):e45.
• [19] FANG S,CHEN L,ZHAO M P.Unimolecular chemically modified DNA fluorescent probe for one-step quantitative measurement of the activity of human apurinic/apyrimidinic endonuclease 1 in biological samples[J].Analytical Chemistry,2015,87(24):11952-11956.
• [20] FAN Z T,ZHAO J,CHAI X,et al.A cooperatively activatable,DNA-based fluorescent reporter for imaging of correlated enzymatic activities[J].Angewandte Chemie,2021,60(27):14887-14891.
• [21] LI X J,XIONG M,HUANG Y F,et al.Simple label-free fluorescence detection of apurinic/apyrimidinic endonuclease 1 activity and its inhibitor using the abasic site-binding fluorophore[J].Analytical Methods,2019,11(6):739-743.
• [22] LV M M,LIU J W,YU R Q,et al.A bipedal DNA nanowalker fueled by catalytic assembly for imaging of base-excision repairing in living cells[J].Chemical Science,2020,11(38):10361-10366.