Zangrossi Stefano, Marabese Mirko, Broggini Massimo, Giordano Rosaria, D'Erasmo Marco, Montelatici Elisa, Intini Daniela, Neri Antonino, Pesce Maurizio, Rebulla Paolo, Lazzari Lorenza
Cell Factory, Center for Transfusion Medicine, Cellular Therapy and Cryobiology, Fondazione Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Via F. Sforza 35, 20122 Milano, Italy.
Stem Cells. 2007 Jul;25(7):1675-80. doi: 10.1634/stemcells.2006-0611. Epub 2007 Mar 22.
The Oct-4 transcription factor, a member of the POU family that is also known as Oct-3 and Oct3/4, is expressed in totipotent embryonic stem cells (ES) and germ cells, and it has a unique role in development and in the determination of pluripotency. ES may have their postnatal counterpart in the adult stem cells, recently described in various mammalian tissues, and Oct-4 expression in putative stem cells purified from adult tissues has been considered a real marker of stemness. In this context, normal mature adult cells would not be expected to show Oct-4 expression. On the contrary, we demonstrated, using reverse transcription-polymerase chain reaction (PCR) (total RNA, Poly A+), real-time PCR, immunoprecipitation, Western blotting, band shift, and immunofluorescence, that human peripheral blood mononuclear cells, genetically stable and mainly terminally differentiated cells with well defined functions and a limited lifespan, express Oct-4. These observations raise the question as to whether the role of Oct-4 as a marker of pluripotency should be challenged. Our findings suggest that the presence of Oct-4 is not sufficient to define a cell as pluripotent, and that additional measures should be used to avoid misleading results in the case of an embryonic-specific gene with a large number of pseudogenes that may contribute to false identification of Oct-4 in adult stem cells. These unexpected findings may provide new insights into the role of Oct-4 in fully differentiated cells. Disclosure of potential conflicts of interest is found at the end of this article.
八聚体结合转录因子4(Oct-4)是POU家族的成员之一,也被称为Oct-3和Oct3/4,在全能胚胎干细胞(ES)和生殖细胞中表达,并且在发育和多能性的决定中具有独特作用。ES在成体干细胞中可能有其出生后的对应物,最近在各种哺乳动物组织中都有描述,并且从成年组织中纯化的假定干细胞中Oct-4的表达被认为是干性的真正标志。在这种情况下,正常成熟的成年细胞预计不会显示Oct-4表达。相反,我们使用逆转录-聚合酶链反应(PCR)(总RNA、Poly A+)、实时PCR、免疫沉淀、蛋白质印迹、条带迁移和免疫荧光证明,人类外周血单个核细胞,这种基因稳定且主要为终末分化、功能明确且寿命有限的细胞,表达Oct-4。这些观察结果提出了一个问题,即Oct-4作为多能性标志的作用是否应该受到挑战。我们的研究结果表明,Oct-4的存在不足以将一个细胞定义为多能性细胞,并且在存在大量假基因的胚胎特异性基因的情况下,应该采用其他措施来避免误导性结果,因为这些假基因可能导致在成体干细胞中对Oct-4的错误鉴定。这些意外发现可能为Oct-4在完全分化细胞中的作用提供新的见解。潜在利益冲突的披露见本文末尾。