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MULTIPLEXED MOLECULAR PROFILING OF PROSTATE CANCER SPECIMENS USING SEMICONDUCTOR QUANTUM DOT BIOCONJUGATES

机译:半导体量子点生物缀合物对前列腺癌标本的多重分子谱分析

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Quantum dots (QDs) are light emitting semi-conductor nanocrystals with novel optical properties including superior photostability, narrow emission spectra with continuous excitation spectra. These properties make QDs especially suitable for multiplexed fluorescent labeling, live cell imaging, and in vivo animal imaging. The multiplexing potential has been recognized but real applications of biological/clinical significance are few. In this study, we used quantum dots to study epithelial mesenchymal transition (EMT), an important process involved in the bone metastasis of prostate cancer.Two prostate cancer cells lines with distinct molecular profiles, representing the two ends of the EMT process, were selected for this study. Four EMT-related biomarkers including E-cadherin, N-cadherin, Vimentin, and RANKL were stained with QD-antibody conjugates with elongation factor 1 alpha as the internal control. Morphological information of the QD-stained cells was obtained by digital-color imaging and quantitative information obtained by spectra analysis using a spectrometer. Two types of analysis were performed: abundance of each biomarker in the same cell line relative to the internal control; and the relative abundance of these markers between the two cell lines. Our results demonstrate the feasibility of QDs for multiplexed profiling of FFPE cells/tissue of clinical significance; however, the standardization and quantification still awaits optimization.
机译:量子点(QD)是具有新颖光学特性的发光半导体纳米晶体,这些光学特性包括出色的光稳定性,具有连续激发光谱的窄发射光谱。这些特性使量子点特别适合于多重荧光标记,活细胞成像和体内动物成像。已经认识到多路复用潜力,但是生物学/临床意义的实际应用很少。在这项研究中,我们使用量子点研究了上皮间质转化(EMT),这是涉及前列腺癌骨转移的重要过程。选择了两种具有不同分子谱的前列腺癌细胞系,分别代表了EMT过程的两端这项研究。包括E-钙粘着蛋白,N-钙粘着蛋白,波形蛋白和RANKL在内的4种与EMT相关的生物标记物均以QD抗体偶联物染色,其中延伸因子1α为内部对照。通过数字彩色成像获得QD染色细胞的形态学信息,并通过使用分光计的光谱分析获得定量信息。进行了两种类型的分析:相对于内部对照,同一细胞系中每种生物标志物的丰度;以及两个细胞系之间这些标记的相对丰度。我们的结果证明了QDs对FFPE细胞/组织进行多重分析的临床意义。但是,标准化和量化仍在等待优化。

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