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一种用于检测 SKBR-3 细胞(转移性乳腺癌细胞系)的高灵敏电化学发光细胞传感器:早期和精确诊断的建设性阶段。

A highly sensitive electrochemiluminescence cytosensor for detection of SKBR-3 cells as metastatic breast cancer cell line: A constructive phase in early and precise diagnosis.

机构信息

Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.

Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Biosens Bioelectron. 2021 Apr 15;178:113023. doi: 10.1016/j.bios.2021.113023. Epub 2021 Jan 21.

Abstract

Ultrasensitive monitoring of cancer cells, especially metastatic ones, has a great interest in human medicine. Despite the early diagnosis of diseases, there is an essential need for any prediction in the severity of side effects for therapeutic outcomes like metastasis. Therefore, the inhibition of cancer cells metastasis to other organs is of utmost importance for cancer suffering patients. In this regard, we developed an electrochemiluminescence (ECL)-based cytosensor for the quantification of metastatic breast cancer cells, namely SKBR-3. Silica-based mesoporous materials have a great potential for application in ECL biosensors due to their high loading capacity and mechanical strength. Herein, a silica-based electrode was prepared via in situ electrosyntheses of mesoporous silica as an environmentally friendly approach. In this protocol, luminol (as luminophore) was combined with chitosan (as attachment biomolecule) to produce a stable lumino-composite film on the electrode surface. At the optimum experimental conditions, the lower limit of quantitation (LLOQ) and linear dynamic range (LDR) were obtained as 20 cells/mL and 20 to 2000 cells/mL, individually. The specificity was desirably examined in the presence of other breast cancer cell lines such as MCF-7 and MDA-MB-231, as a model of early-stage and invasive phases of breast cancer cells. The repeatability was successfully examined for five repetitive measurements and the acceptable relative standard deviation (RSD) was calculated as about 1.6% for 500 cells/mL. As a proof of concept, the presented cytosensor has a high ability to use in clinical laboratories for the detection and separation of metastatic cells via the combination with microfluidic systems.

摘要

超敏监测癌细胞,特别是转移性癌细胞,在人类医学中具有重要意义。尽管疾病的早期诊断很重要,但对于治疗结果(如转移)的副作用严重程度的任何预测都非常必要。因此,抑制癌细胞向其他器官转移对于癌症患者至关重要。在这方面,我们开发了一种基于电化学发光(ECL)的细胞传感器,用于定量检测转移性乳腺癌细胞,即 SKBR-3。基于硅的介孔材料由于其高负载能力和机械强度,在 ECL 生物传感器中有很大的应用潜力。在此,通过原位电合成介孔硅制备了基于硅的电极,这是一种环保的方法。在本方案中,将鲁米诺(作为发光体)与壳聚糖(作为附着生物分子)结合,在电极表面产生稳定的发光复合膜。在最佳实验条件下,分别获得了 20 个细胞/mL 的定量下限(LLOQ)和 20 到 2000 个细胞/mL 的线性动态范围(LDR)。在存在其他乳腺癌细胞系(如 MCF-7 和 MDA-MB-231)的情况下,对特异性进行了理想的检查,这些细胞系是乳腺癌细胞早期和侵袭阶段的模型。对五个重复测量的重复性进行了成功检查,可接受的相对标准偏差(RSD)约为 500 个细胞/mL 时的 1.6%。作为概念验证,该细胞传感器具有通过与微流控系统结合用于临床实验室中检测和分离转移性细胞的高能力。

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