University Department of Growth and Reproduction, Section GR5064, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark.
Mol Hum Reprod. 2010 Nov;16(11):835-45. doi: 10.1093/molehr/gaq008. Epub 2010 Jan 31.
The transcription factor OCT4 plays a crucial role in the earliest differentiation of the mammalian embryo and in self-renewal of embryonic stem cells. However, it remains controversial whether this gene is also expressed in somatic tissues. Here, we use a combination of RT-PCR on whole and microdissected tissues, in situ hybridization, immunohistochemistry and western blotting to show that OCT4 and SOX2 together with downstream targets, UTF1 and REX1/ZFP42, are expressed in the human male urogenital tract. We further support these results by the analysis of DNA methylation of a region in the OCT4 promoter. In culture, human primary epididymal cells formed spheres that continued to express the investigated genes for at least 20 days. Transcriptomic analysis of cultured cells showed up-regulation of CD29, CD44 and CD133 that are normally associated with sphere-forming cancer stem cells. Furthermore, stimulation with retinoic acid resulted in down-regulation of OCT4 expression, however, without multilineage differentiation. Our results show that OCT4 and associated genes are expressed in somatic epithelial cells from the urogenital tract and that these cells can form spheres, a general marker of stem cell behaviour.
转录因子 OCT4 在哺乳动物胚胎的最早分化和胚胎干细胞的自我更新中起着至关重要的作用。然而,关于该基因是否也在体组织中表达,仍存在争议。在这里,我们使用 RT-PCR 对整体和微切割组织、原位杂交、免疫组织化学和 Western blot 进行组合分析,结果表明 OCT4 和 SOX2 与下游靶标 UTF1 和 REX1/ZFP42 一起在人类男性泌尿生殖系统中表达。我们还通过分析 OCT4 启动子区域的 DNA 甲基化来支持这些结果。在培养中,人原发性附睾细胞形成球体,至少持续表达研究基因 20 天。培养细胞的转录组分析显示,通常与球体形成的癌症干细胞相关的 CD29、CD44 和 CD133 的表达上调。此外,用维甲酸刺激导致 OCT4 表达下调,但没有多谱系分化。我们的结果表明,OCT4 和相关基因在泌尿生殖系统的体上皮细胞中表达,这些细胞可以形成球体,这是干细胞行为的一个普遍标志。