端粒酶活性检测试纸构建与肿瘤细胞可视化判定

引用本文: 张瑞雪, 陈文静, 姜玮, 徐晓文. 端粒酶活性检测试纸构建与肿瘤细胞可视化判定[J]. 分析化学, 2021, 49(5): 702-709. doi: 10.19756/j.issn.0253-3820.201765 shu

Citation: ZHANG Rui-Xue, CHEN Wen-Jing, JIANG Wei, XU Xiao-Wen. Construction of Telomerase Activity Test Strip and Visual Determination of Cancerous Cells[J]. Chinese Journal of Analytical Chemistry, 2021, 49(5): 702-709. doi: 10.19756/j.issn.0253-3820.201765 shu

端粒酶活性检测试纸构建与肿瘤细胞可视化判定

山东大学化学与化工学院, 济南 250100

通讯作者: 徐晓文,E-mail:xuxw@sdu.edu.cn

基金项目:
国家自然科学基金项目（No.21705094）、电分析化学国家重点实验室开放课题项目（No.SKLEAC202004）和山东大学青年学者未来计划项目（No.2018WLJH50）资助。

摘要: 端粒酶是将端粒重复序列添加到染色体末端端粒上的核糖核蛋白，可补偿细胞分裂过程中的端粒缩短，维持细胞持续增殖。端粒酶活性在肿瘤细胞中过表达，在正常体细胞中被抑制，是癌症诊断和预后评估的重要标志物。本研究构建了一种检测试纸，用于可视化分析端粒酶活性和判定肿瘤细胞。将样品垫、喷涂DNA功能化金纳米粒子的结合垫、含有测试区和对照区的硝酸纤维素膜和吸收垫共同组装在底板上，构成检测试纸。在端粒酶的催化作用下，底物序列发生链延伸，然后被滴加在样品垫上。溶液依靠毛细作用在试纸上流动，水化结合垫上的DNA功能化金纳米粒子。在流动过程中，端粒酶延伸产物、测试区中捕获的DNA、金纳米粒子上修饰的DNA杂交形成三明治式结构，使金纳米粒子富集在测试区，T线显示红色。同时，DNA功能化金纳米粒子继续流动，与对照区中捕获DNA杂交，C线显示红色，以验证试纸的有效性。通过响应细胞裂解液中的端粒酶，本方法可通过观察T线显色的方式对肿瘤细胞和正常细胞进行简便、快速的可视化区分。

关键词:

English

Construction of Telomerase Activity Test Strip and Visual Determination of Cancerous Cells

School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China

Corresponding author: XU Xiao-Wen, xuxw@sdu.edu.cn

Received Date: 17 December 2020
Revised Date: 17 January 2021
Fund Project:
Supported by the National Natural Science Foundation of China (No.21705094), the Open Funds of the State Key Laboratory of Electroanalytical Chemistry (No.SKLEAC202004), and the Young Scholars Program of Shandong University (No.2018WLJH50).

Abstract: Telomerase is a ribonucleoprotein that can catalyze the addition of telomerase repeats onto the end of telomere, compensating for the telomere shortening during cell division and maintaining the continuous cell proliferation. Telomerase activity is overexpressed in cancerous cells but inhibited in normal somatic cells, and thus it is regarded as an important biomarker for cancer diagnosis and prognosis evaluation. In this work, a test strip is constructed for visual detection of telomerase activity and determination of cancerous cells. The test strip is constituted of sample pad, conjugate pad that is sprayed with DNA-functionalized gold nanoparticles, nitrocellulose membrane that contains test zone and control zone, and absorbent pad, which are all assembled onto the backing plate. Under the action of telomerase, the substrate sequence gets extended, which are then dropped onto the sample pad. The solution migrates by capillary action and rehydrates the DNA-functionalized AuNPs on the conjugate pad. Subsequently, the telomeric products hybridize with the capture DNA in the test zone and the modified DNA on AuNPs in a sandwich manner, causing the accumulation of AuNPs on the test zone and the color development of T-line. Meanwhile, the DNA-functionalized AuNPs continue flowing and then hybridize with the capture DNA on the control zone, causing the color development of C-line and confirming the validity of the test strip. Through responding to the telomerase from cell extracts, the method can achieve the visual differentiation of cancerous cells from normal cells in a convenient and rapid way by observation of color development of T-line.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

端粒酶活性检测试纸构建与肿瘤细胞可视化判定

通讯作者: 徐晓文,E-mail:xuxw@sdu.edu.cn

English

Construction of Telomerase Activity Test Strip and Visual Determination of Cancerous Cells

Corresponding author: XU Xiao-Wen, xuxw@sdu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

端粒酶活性检测试纸构建与肿瘤细胞可视化判定

通讯作者: 徐晓文,E-mail:xuxw@sdu.edu.cn

English

Construction of Telomerase Activity Test Strip and Visual Determination of Cancerous Cells

Corresponding author: XU Xiao-Wen, xuxw@sdu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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