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ANALYTICAL CHEMISTRY · 2017 · IF 6.32

Sensitive and Label-Free Fluorescent Detection of Transcription Factors Based on DNA-Ag Nanoclusters Molecular Beacons and Exonuclease III-Assisted Signal Amplification

Bingzhi Li,Lei Xu,Yue Chen,Wanying Zhu,Xin Shen,Chunhong Zhu,Jieping Luo,Xiaoxu Li,Junli Hong,Xuemin Zhou

Transcription factors (TFs) regulate gene expression by binding to regulatory regions, and their dysregulation is involved in numerous diseases. Thus, they are therapeutic targets and potential diagnostic markers. However, widely used methods for TFs detection are either cumbersome or costly. Herein, we first applied DNA-Ag nanoclusters molecular beacons (AgMBs) in TFs analysis and designed an assay based on the switchable fluorescence of AgMBs. In the absence of TFs, a single-stranded DNA functioned as a reporter is released from a double-stranded DNA probe (referred as dsTFs probe) under exonuclease III (Exo III) digestion. Then, the reporter triggers downstream Exo III-assisted signal amplification by continuously consuming the guanine-rich enhancer sequences in AgMBs, resulting in significant fluorescent decrease eventually. Conversely, the presence of TFs protects the dsTFs probe from digestion and blocks the downstream reaction to keep a highly fluorescent state. To testify this rationale, we utilized nuclear factor-kappa B p50 (NF-κB p50) as a model TFs. Owing to the amplification strategy, this method exhibited high sensitivity toward NF-κB p50 with a limit of detection of 10 pM, and a broad linear range from 30 pM to 1.5 nM. Furthermore, this method could detect multiple TFs in human colon cancer DLD-1 cells and reflect the variation in their cellular levels after stimulation. Finally, by conducting an inhibition assay we revealed the potential of this method for screening TFs-targeted drugs and calculating the IC50 of corresponding inhibitors.

CAS / MDL: 16940-66-2 7761-88-8
研究领域: 生物化学 · 分子生物学 · 生物技术 · 癌症研究 · 药物发现
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ANALYST · 2014 ·Vol. 139 ·期 19 ·4917-4923 · IF 3.906

A novel aptasensor based on MUC-1 conjugated CNSs for ultrasensitive detection of tumor cells

Hongmei Cao,Daixin Ye,Qianqian Zhao,Juan Luo,Song Zhang,Jilie Kong

A novel strategy for the quantitative determination of human colon cancer DLD-1 cells utilizing an electrochemical aptasensor was developed by effective surface recognition between Mucin 1 glycoprotein over-expressed on the cell membrane and MUC-1 aptamer (MUC-1) bound on carbon nanospheres (CNSs). An MTT assay revealed that the as-prepared CNSs by green route exhibited satisfactory biocompatibility for cell viability, providing a suitable platform for the cell adhesion study. Furthermore, using CNSs as a sensing layer accelerated electron transfer and provided a highly stable matrix for the convenient conjugation of target MUC-1 aptamer, considerably amplifying the electrochemical signals. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were applied to assess the optimal conditions and detection performance of the as-fabricated aptasensor. The attachment of colon cancer DLD-1 cells onto the MUC-1 aptamer immobilized CNSs led to increased EIS responses, which changed linearly in cell concentration ranging from 1.25 × 102 to 1.25 × 106 cells per mL with a lower detection limit of 40 cells per mL. With this method, colon cancer DLD-1 cells can be easily distinguished from normal cells, Human astrocytes 1800. The novel aptasensor revealed high specificity to DLD-1 cells. Furthermore, the aptasensor described here showed good reproducibility and high stability because of the CNSs of high stability and biocompatibility. The proposed protocols are a promising technique for the early monitoring of human colon cancer, and might have potential clinical applications such as cancer diagnosis, drug screening.

CAS / MDL: 25952-53-8 6066-82-6
研究领域: 生物传感器 · 电化学 · 癌症诊断 · 纳米技术 · 分析化学
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY · 2022 · IF 5.895

Inhibition of the Occurrence and Development of Inflammation-Related Colorectal Cancer by Fucoidan Extracted from Sargassum fusiforme

Xiang Li,Shijun Xin,Xiaoqun Zheng,Liqin Lou,Shiqing Ye,Shengkai Li,Qilong Wu,Qingyong Ding,Ling Ji,Chunrong Nan,Yongliang Lou

Fucoidan has many biological activities, including the inhibitory effect on the development of various cancer types. This study showed that lipopolysaccharide-induced inflammation in FHC cells (normal human colonic epithelial cells) could be reversed using fucoidan at different concentrations. The fucoidan-induced anti-inflammatory effect was also confirmed through in vivo experiments in mice. Compared to the mice of the model group, the ratio of Firmicutes/Bacteroidetes in feces increased and the diversity of gut microbial composition was restored in mice after fucoidan intervention. In colorectal cancer (CRC) cells DLD-1 and SW480, fucoidan inhibited cell proliferation and promoted cell apoptosis. It also blocked the cell cycle of DLD-1 and SW480 at the G0/G1 phase. The animal model of inflammation-related CRC showed that the incidence of tumors in mice was significantly reduced by fucoidan intervention. Furthermore, the administration of fucoidan decreased the expression levels of inflammatory factors such as TNF-α IL-6 and IL-1β in the colonic tissues. Therefore, fucoidan can effectively prevent the development of colitis-associated CRC.

CAS / MDL: 9011-18-1
研究领域: 癌症生物学 · 免疫学 · 胃肠病学 · 微生物学 · 药理学
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