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首页> 外文期刊>The Analyst >Characterization of microRNA-125b expression in MCF7 breast cancer cells by ATR-FTIR spectroscopy
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Characterization of microRNA-125b expression in MCF7 breast cancer cells by ATR-FTIR spectroscopy

机译：通过ATR-FTIR光谱表征MCF7乳腺癌细胞中microRNA-125b的表达

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MicroRNAs (miRNAs), are u000222 nucleotides long, non-coding RNAs that control gene expressionnpost-transcriptionally by binding to their target mRNA’s 30UTRs (untranslated regions). Due to theirnroles in various important regulatory processes and pathways, miRNAs have been implicated in diseasenmechanisms such as tumorigenesis when their expression is deregulated. To date, a significant numbernof miRNAs and their target messenger RNAs (mRNAs) have been identified and verified. It is generallynaccepted that miRNAs can potentially bind to many mRNAs, which brings the requirement ofnvalidation of these interactions. While understanding that such individual interactions is crucial tondelineate the role of a specific miRNA, we took a holistic approach and analyzed global changes in thencell due to expression of a miRNA in a model cell line system. Our model consisted of MCF7 cellsnstably transfected with miR-125b (MCF7-125b) and empty vector (MCF7-EV). MiR-125b is one of thenknown down-regulated miRNAs in breast cancers. In this study we examined the global structuralnchanges in MCF7 cells lacking and expressing miR-125b by Attenuated Total Reflectance FouriernTransform Infrared (ATR-FTIR) Spectroscopy and investigated the dynamic changes by morensensitive spin-labelling Electron Spin Resonance (ESR) spectroscopy. Our results revealed less RNA,nprotein, lipid, and glycogen content in MCF7-125b compared to MCF7-EV cells. Membrane fluiditynand proliferation rate were shown to be lower in MCF7-125b cells. Based on these changes, MCF7-n125b and MCF7-EV cells were discriminated successfully by cluster analysis. Here, we provide a novelnmeans to understand the global effects of miRNAs in cells. Potential applications of this approach arennot only limited to research purposes. Such a strategy is also promising to pioneer the development ofnfuture diagnostic tools for deregulated miRNA expression in patient samples.
机译：MicroRNA（miRNA）长u000222核苷酸，是非编码RNA，通过与靶标mRNA的30UTR（非翻译区）结合来转录后控制基因表达。由于它们在各种重要的调控过程和途径中的作用，miRNA的表达失调已与疾病机制如肿瘤发生有关。迄今为止，已经鉴定并验证了大量的miRNA及其靶信使RNA（mRNA）。人们普遍接受miRNA可能与许多mRNA结合，这带来了验证这些相互作用的要求。虽然了解到此类个体相互作用至关重要，但它却说明了特定miRNA的作用，但我们采用了一种整体方法，并分析了由于miRNA在模型细胞系系统中表达而导致thencell的整体变化。我们的模型由稳定转染了miR-125b（MCF7-125b）和空载体（MCF7-EV）的MCF7细胞组成。 MiR-125b是乳腺癌中当时已知的下调miRNA之一。在这项研究中，我们通过衰减全反射傅立叶变换红外（ATR-FTIR）光谱技术检查了缺少和表达miR-125b的MCF7细胞的整体结构变化，并通过更敏感的自旋标记电子自旋共振（ESR）光谱研究了动态变化。我们的结果显示，与MCF7-EV细胞相比，MCF7-125b中的RNA，n蛋白，脂质和糖原含量更少。 MCF7-125b细胞膜流动性和增殖率较低。基于这些变化，通过聚类分析成功地区分了MCF7-n125b和MCF7-EV细胞。在这里，我们提供了一种新颖的方法来了解miRNA在细胞中的整体作用。这种方法的潜在应用不仅限于研究目的。这种策略也有望开创用于诊断患者样品中miRNA表达失调的未来诊断工具的先河。

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  • 来源
    《The Analyst》 |2010年第12期|p.3094-3102|共9页
  • 作者

    Nihal Simsek Ozek Serkan Tuna A. Elif Erson-Bensan and Feride Severcan;

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    Middle East Technical University, Department of Biological Sciences,06530 Ankara, Turkey. E-mail: feride@metu.edu.tr;

    Fax: +90 312 21076 79;

    Tel: +90 312 210 51 66† This article is part of a themed issue on Optical Diagnosis. This issueincludes work presented at SPEC 2010 Shedding Light on Disease:Optical Diagnosis for the New Millennium, which was held inManchester, UK June 26th–July 1st 2010.‡ Electronic supplementary information (ESI) available: The FTIRspectra of twenty independent samples of MCF7-EV and MCF7-125bcells, together with the representative FTIR spectra of native and PBSsubtracted MCF7-EV cells and second derivative spectra of RNA andDNA purified from MCF7 cells. See DOI: 10.1039/c0an00543f;

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