Maragkoudaki Maria, Vaiopoulou Anna, Theodoropoulos George E, Legaki Evangelia, Sechi Leonardo A, Karamanolis George, Zografos George, Gazouli Maria
First Department of Pediatrics, Athens University Medical School, "Aghia Sophia" Children's Hospital, Athens, Greece.
Laboratory of Biology, Department of Basic Medical Sciences, School of Medicine, University of Athens, Michalakopoulou 176, 11527 Athens, Greece.
Gut Pathog. 2015 Oct 1;7:25. doi: 10.1186/s13099-015-0073-1. eCollection 2015.
Developmentally early cells are mobilized into peripheral blood in Crohn's disease (CD) patients. OCT4, is considered to be important in sustaining the pluripotency of stem cells. OCT4 splicing variants are differentially expressed in pluripotent and non-pluripotent cells. Our study aims to investigate the expression pattern of OCT4 variants and SOX-2, an essential factor implicated in self-renewal and pluripotency, in tissue and blood samples from patients with IBD.
Peripheral blood and tissue samples were collected from patients with active CD and ulcerative colitis (UC), and from healthy individuals. OCT4 expression was documented by Western blot, immunohistochemistry and by reverse transcription-real-time PCR. OCT4 isoform determination was documented using specific primers. SOX-2 expression levels were also evaluated.
OCT4 protein levels were significantly higher in CD tissue samples than in CD blood samples, and in UC tissue samples. OCT4 protein was localized mainly in the cytosol. In all samples, only the OCT4 pseudogenes and the OCT4B1 variant were detected. OCT4B1 expression levels were elevated in both tissue and blood samples from CD and UC cases compared to healthy controls. In CD patients only SOX-2 mRNA levels were found slightly increased compared to healthy controls.
Our results suggest that OCT4 is expressed in patients with IBD. Furthermore, we found the presence of the OCT4B1 isoform in IBD in both tissue and blood samples. Our results have shown, that developmentally early cells might be mobilized into peripheral blood as result of tissue damage, indicating a possible role of these cells in repair of injured intestinal tract.
在克罗恩病（CD）患者中，发育早期的细胞会被动员到外周血中。OCT4被认为在维持干细胞的多能性方面很重要。OCT4剪接变体在多能和非多能细胞中差异表达。我们的研究旨在调查OCT4变体和SOX-2（一种与自我更新和多能性有关的关键因子）在炎症性肠病（IBD）患者的组织和血液样本中的表达模式。
收集活动性CD和溃疡性结肠炎（UC）患者以及健康个体的外周血和组织样本。通过蛋白质免疫印迹、免疫组织化学和逆转录实时PCR记录OCT4的表达。使用特异性引物记录OCT4异构体的测定。还评估了SOX-2的表达水平。
CD组织样本中的OCT4蛋白水平显著高于CD血液样本和UC组织样本。OCT4蛋白主要定位于细胞质中。在所有样本中，仅检测到OCT4假基因和OCT4B1变体。与健康对照相比，CD和UC病例的组织和血液样本中OCT4B1的表达水平均升高。在CD患者中，仅发现SOX-2 mRNA水平与健康对照相比略有增加。
我们的结果表明OCT4在IBD患者中表达。此外，我们发现在IBD的组织和血液样本中均存在OCT4B1异构体。我们的结果表明，发育早期的细胞可能由于组织损伤而被动员到外周血中，表明这些细胞在受损肠道修复中可能发挥作用。
