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Specific detection of OCT4 isoforms in inflammatory bowel disease.

Maragkoudaki M, Vaiopoulou A, Theodoropoulos GE, Legaki E, Sechi LA, Karamanolis G, Zografos G, Gazouli M - Gut Pathog (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: First Department of Pediatrics, Athens University Medical School, "Aghia Sophia" Children's Hospital, Athens, Greece.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of representative quantitative real-time reverse transcription PCR analysis for OCT4 isoform identification. The primers and the methodology used was performed as previously described [10, 14, 16]. Samples 1, 2, 3 were from tissues of healthy, CD and UC, respectively. Samples 4, 5, and 6 were from blood of healthy, CD and UC, respectively
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Fig3: Schematic representation of representative quantitative real-time reverse transcription PCR analysis for OCT4 isoform identification. The primers and the methodology used was performed as previously described [10, 14, 16]. Samples 1, 2, 3 were from tissues of healthy, CD and UC, respectively. Samples 4, 5, and 6 were from blood of healthy, CD and UC, respectively

Mentions: In order to reliably discriminate isoforms OCT4A from OCT4B, OCT4B1 and all the known pseudogenes we used the RT-PCR method followed by restriction fragment analysis and OCT4B1 specific primers as previously described [10, 14, 16]. As indicated in Fig. 3 in all cases only the OCT4 pseudogenes and the OCT4B1 variant were detected. The expression pattern of OCT4B1 variant in CD, UC and healthy tissues and blood samples was further examined. As indicated in Fig. 4, OCT4B1 expression levels were elevated in both tissue and blood samples from CD and UC cases compared to healthy controls. Particularly, in blood samples OCT4B1 was expressed 6.95 ± 1.59-fold greater in CD and 3.55 ± 0.57-fold greater in UC compared with healthy controls. Similar results were obtained in tissues samples, also (5.45 ± 1.12-fold greater in CD and 2.74 ± 0.53-fold greater in UC, respectively). The OCT4B1 mRNA levels were higher in blood samples compared to tissue samples from CD patients. The samples from CD patients expressed higher levels of OCT4B1 mRNA compared to respective samples from the UC patients. The mRNA levels of SOX-2 were found slightly increased compared to healthy controls, in both blood (0.91 ± 0.17-fold) and tissue samples (0.84 ± 0.14-fold) of CD patients only.Fig. 3


Specific detection of OCT4 isoforms in inflammatory bowel disease.

Maragkoudaki M, Vaiopoulou A, Theodoropoulos GE, Legaki E, Sechi LA, Karamanolis G, Zografos G, Gazouli M - Gut Pathog (2015)

Schematic representation of representative quantitative real-time reverse transcription PCR analysis for OCT4 isoform identification. The primers and the methodology used was performed as previously described [10, 14, 16]. Samples 1, 2, 3 were from tissues of healthy, CD and UC, respectively. Samples 4, 5, and 6 were from blood of healthy, CD and UC, respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4591585&req=5

Fig3: Schematic representation of representative quantitative real-time reverse transcription PCR analysis for OCT4 isoform identification. The primers and the methodology used was performed as previously described [10, 14, 16]. Samples 1, 2, 3 were from tissues of healthy, CD and UC, respectively. Samples 4, 5, and 6 were from blood of healthy, CD and UC, respectively
Mentions: In order to reliably discriminate isoforms OCT4A from OCT4B, OCT4B1 and all the known pseudogenes we used the RT-PCR method followed by restriction fragment analysis and OCT4B1 specific primers as previously described [10, 14, 16]. As indicated in Fig. 3 in all cases only the OCT4 pseudogenes and the OCT4B1 variant were detected. The expression pattern of OCT4B1 variant in CD, UC and healthy tissues and blood samples was further examined. As indicated in Fig. 4, OCT4B1 expression levels were elevated in both tissue and blood samples from CD and UC cases compared to healthy controls. Particularly, in blood samples OCT4B1 was expressed 6.95 ± 1.59-fold greater in CD and 3.55 ± 0.57-fold greater in UC compared with healthy controls. Similar results were obtained in tissues samples, also (5.45 ± 1.12-fold greater in CD and 2.74 ± 0.53-fold greater in UC, respectively). The OCT4B1 mRNA levels were higher in blood samples compared to tissue samples from CD patients. The samples from CD patients expressed higher levels of OCT4B1 mRNA compared to respective samples from the UC patients. The mRNA levels of SOX-2 were found slightly increased compared to healthy controls, in both blood (0.91 ± 0.17-fold) and tissue samples (0.84 ± 0.14-fold) of CD patients only.Fig. 3

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: First Department of Pediatrics, Athens University Medical School, "Aghia Sophia" Children's Hospital, Athens, Greece.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

No MeSH data available.


Related in: MedlinePlus