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【资讯翻译】Gene Expression Profiling of Circulating Tumor Cells in Bre...

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发表于 2015-1-7 10:17:03 | 显示全部楼层 |阅读模式
Gene Expression Profiling of Circulating Tumor Cells in Breast Cancer

BACKGROUND: Determining the transcriptional profile of circulating tumor cells (CTCs) may allow the acquisition of clinically relevant information while overcoming tumor heterogeneity-related biases associated with use of tissue samples for biomarker assessment. However, such molecular characterization is challenging because CTCs are rare and outnumbered by blood cells.
METHODS: Here, we describe a technical protocol to measure the expression of >29 000 genes in CTCs captured from whole blood with magnetic beads linked with antibodies against epithelial cell adhesion molecule (EpCAM) and the carcinoma-associated mucin, MUC1, designed to be used for CTC characterization in clinical samples. Low numbers of cells (5–200) from the MCF7 and MDA-MB-468 breast cancer cell lines were spiked in healthy donor blood samples and isolated with the AdnaTest EMT-1/Stem CellSelect kit. Gene expression profiles (GEPs) were obtained with the WG-DASL HT assay and compared with GEPs obtained from RNA isolated from cultured cell lines and unspiked samples.
RESULTS: GEPs from samples containing 25 or more spiked cells correlated (r = 0.95) with cognate 100-ng RNA input samples, clustered separately from blood control samples, and allowed MCF7 and MDA-MB-468 cells to be distinguished. GEPs with comparable technical quality were also obtained in a preliminary series of clinical samples.
CONCLUSIONS: Our approach allows technically reliable GEPs to be obtained from isolated CTCs for the acquisition of biologically useful information. It is reproducible and suitable for application in prospective studies to assess the clinical utility of CTC GEPs, provided that >25 CTCs can be isolated.

信源地址:http://www.clinchem.org/content/61/1/278.abstract

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发表于 2015-1-7 12:08:10 | 显示全部楼层
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发表于 2015-1-7 21:08:00 | 显示全部楼层
乳腺癌循环肿肿瘤细胞的基因表达谱

确定循环肿瘤细胞的转录谱可以获得临床相关信息,并且可能克服使用组织样品进行生物标记物评估所带来的肿瘤细胞异质性相关误差。然而,该项分子技术具有很高的挑战性,因为在血液中的循环肿瘤细胞非常少,其数量远远低于红细胞。

在最新一期的《Clinical Chemistry》杂志中Cappelletti V小组描述了一项技术操作说明,他们通过使用耦合有抗表面粘附分子(EpCAM)以及肿瘤相关黏连蛋白(MUC1) 的抗体的磁珠去从临床样品中分离出肿瘤循环细胞,去检测从全血中分离出来的肿瘤循环细胞中大于29000个基因表达情况,。从MCF7以及MDA-MB-468肿瘤细胞系中获得的数量较少的细胞(5-200)被添加健康的供体血液样本并且使用AdnaTest EMT-1/Stem Cell Select 试剂盒进行分选。 基因表达谱通过WG-DASL HT 方法获得并且同从培养细胞系中以及未添加样本中获得的RNA基因表达谱相比较。

结果显示:从包含25个或者更多的循环肿瘤细胞获得的基因表达谱与同源的100ng RNA输入的样本相关(r=0.95),分别地从血液对照样本中积簇,并且能够区分MCF7以及MDA-MB-468细胞系。基因表达谱比较技术质量参数同样在前期一系列的临床样本中获得。

Cappelletti V等认为该方法允许从分离出来的循环肿瘤细胞中获得技术可靠的基因表达谱进而获得有用的生物学信息。如果我们能够分离到大于25个肿瘤循环细胞,那么在临床上使用肿瘤循环细胞基因表达谱的测定将是具有很好的前景和应用的。

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 楼主| 发表于 2015-1-9 12:05:32 | 显示全部楼层
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