Abstract
Abstract Several proteins, including Oct-3/4, Sox2, KLF4 and Nanog, play a critical role in the maintenance of pluripotency in stem cells. Overexpression of one or a combination of these factors has been reported in solid tumors from a variety of tissues. The presence of pseudogenes and differentially expressed protein isoforms complicates the accurate detection of the physiologically relevant forms of these proteins in cancer stem cell research. Here, we report the development of MILLIPLEX® MAP multiplex assays for the simultaneous, specific and sensitive detection of the pluripotency markers Oct-3/4, Sox2, KLF4 and Nanog as well as several other stem cell relevant proteins in cellular lysates. Application of the assay to a panel of 25 human cancer cell lines from a variety of organs revealed Sox2 to be the most often expressed protein among these lines. Of the cell lines assayed, only one breast cancer cell line expressed detectable levels of all three transcription factors, suggesting a role for Oct3/4, Sox2 and Nanog in cancer stem cells of this particular cell line. These experiments demonstrate the utility of this pluripotency marker panel for the sensitive analysis of stem cells from a variety of lineages. The expression of the panel analytes was also examined in induced pluripotent stem cells (iPS) and a human embryonic stem cell (hES) line for reference. Taken together we show the successful development of a multiplex immunoassay that can be used to investigate the role of pluripotency marker proteins in stem cells or cancer cell immortalized or primary lines. Citation Format: Frederick W. Wiese, Nikolai Stankiewicz, Ralf Reiners, Tim Warmke, Reeti Maheshwari. Multiplex detection of stem cell pluripotency markers in reference cancer cell lines. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2652. doi:10.1158/1538-7445.AM2013-2652
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