OBJECTIVE: To discuss the meanings of Oct-4's expression in thyroid adenoma, thyroid papillary carcinoma, thyroid follicular carcinoma, and medullary thyroid carcinoma. METHOD: We examined the expression of Oct-4 in 15 thyroid adenoma, 30 thyroid papillary carcinomas, 2 thyroid follicular carcinomas, and 3 medullary thyroid carcinomas using immunofluorescence. RESULT: Oct-4 expression was observed in all the thyroid-related diseases mentioned above. In thyroid papillary carcinomas, the expression of Oct-4 were higher than that in thyroid adenoma, and had no obvious relationship with the patients age, sex, the size and location of tumor and tumor metastasis. CONCLUSION The formation of the thyroid carcinomas may be concerned with the stem cells in thyroid. There are more stem cells in medullary thyroid carcinomas and follicular carcinomas.
Adenocarcinoma, Follicular ; metabolism ; pathology ; Carcinoma, Neuroendocrine ; Carcinoma, Papillary ; metabolism ; pathology ; Humans ; Octamer Transcription Factor-3 ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology

OBJECTIVETo study the expression of OCT4 protein in bladder cancer and its correlation to the clinicopathologic features and prognosis of bladder cancer.

METHODSOCT4 mRNA and protein expression was detected in 5 bladder cancer cell lines (RT-4, Tcc-Sup, KK47, T24, and 5637) and 1 normal bladder cell lines by real-time PCR and Western blotting, respectively. Immunohistochemical analysis was used to detect the expression of OCT4 protein in 46 bladder cancer samples.

RESULTSAll the 5 bladder cancer cell lines expressed detectable levels of OCT4 mRNA and proteins, whereas the normal bladder cell line SV-HUC-1 was negative for OCT4 expression. The clinical bladder cancer tissues showed a high positivity rate of OCT4 expression (76.1%), which was not detected in normal bladder tissues. Specific OCT-4 signals were localized mainly in the nuclei of the cancer cells. The expression rate of OCT4 protein was significantly higher in bladder cancer tissue than in normal bladder epithelium (P<0.05), and showed a positive correlation to the grade of tumor differentiation and metastasis (P<0.05) but not to the patients' age, gender or TNM stage.

CONCLUSIONOCT4 protein expression is associated with tumor differentiation and metastasis in bladder cancer and may play an important role in the early diagnosis and prognostic evaluation of bladder cancer.

Cell Line, Tumor ; Humans ; Neoplasm Metastasis ; Neoplasm Staging ; Octamer Transcription Factor-3 ; metabolism ; Prognosis ; Urinary Bladder Neoplasms ; diagnosis ; metabolism ; pathology

Mesenchymal stem cells (MSCs) are attractive candidates for clinical repair or regeneration of damaged tissues. Oct4 and Sox2, which are essential transcription factors for pluripotency and self-renewal, are naturally expressed in MSCs at low levels in early passages, and their levels gradually decrease as the passage number increases. Therefore, to improve MSC proliferation and stemness, we introduced human Oct4 and Sox2 for conferring higher expansion and differentiation capabilities. The Oct4-IRES-Sox2 vector was transfected into human adipose tissue MSCs (ATMSCs) by liposomal transfection and used directly. Oct4 and Sox2 were successfully transfected into ATMSCs, and we confirmed maintenance of MSC surface markers without alterations in both red fluorescent protein (RFP) (control) and Oct4/Sox2-ATMSCs. Enhanced proliferative activity of Oct4/Sox2-ATMSCs was shown by WST-1 assay, and this result was further confirmed by cell counting using trypan blue exclusion for a long period. In addition, FACs cell cycle analysis showed that there was a reduction in the fraction of Oct4/Sox2-ATMSCs in G1 with a concomitant increase in the fraction of cells in S, compared with RFP-ATMSCs. Increased levels of cyclin D1 were also seen in Oct4/Sox2-ATMSCs, indicating acceleration in the transition of cells from G1 to S phase. Furthermore, Oct4/Sox2-overexpressing ATMSCs showed higher differentiation abilities for adipocytes or osteoblasts than controls. The markers of adipogenic or osteogenic differentiation were also upregulated by Oct4/Sox2 overexpression. The improvement in cell proliferation and differentiation using Oct4/Sox2 expression in ATMSCs may be a useful method for expanding the population and increasing the stemness of ATMSCs.
Adipose Tissue/cytology ; *Cell Differentiation ; *Cell Proliferation ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells/cytology/*metabolism/physiology ; Octamer Transcription Factor-3/genetics/*metabolism ; SOXB1 Transcription Factors/genetics/*metabolism

Animals ; Astrocytes ; cytology ; Cell Differentiation ; Cellular Reprogramming ; Induced Pluripotent Stem Cells ; cytology ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Mice ; Neurons ; cytology ; Octamer Transcription Factor-3 ; genetics ; metabolism

OBJECTIVETo investigate the methylation Oct4 in orientation induced differentiation in bone marrow mesenchymal stem cells

METHODSMice BMSCs were isolated and purified from bone marrow by adherent culture,and then identified by morphology and immunocytochemistry.Mouse osteoblastic cells were cultured by bone fragments inoculation,and then identified by alkaline phosphatase(AKP)staining and alizarin red staining.BMSCs were induced to differentiate into osteoblasts in vitro. Indirect immunofluorescence staining and reverse transcription polymerase chain reaction(RT PCR)were used to detect the expressions of Oct4 in BMSCs before and after induction.The methylation status of Oct4 gene in mouse BMSCs was explored by a methylation specific PCR before and after induction

RESULTSThe isolated mice BMSCs massively proliferated in vitro and formed cell colones with uniform morphology.Positive expressions of CD29,cKit,and CD44 and negative expression of CD34 were found in the isolated cells.After 10 days[DK]'[DK] induction,both AKP and the alizarin red were positive in cells and osteoblastic cells isolated from mice skull bones.The indirect immunoinfluorescence staining and RT-PCR also showed that the Oct4 expression in the directed differentiation of mouse BMSCs was down-regulated.The CpG island of Otc4 gene promoter in mouse BMSCs became methylated during the induced differentiation.

CONCLUSIONSMice BMSCs and osteoblasts were successfully cultured in vitro in this studyOct4 may be involved in the maintenance of adult stem cell pluripotency.The down regulated expression of Oct4 gene in mouse BMSCs during the directed differentiation may contribute to the methylation of CpG island in Otc4 gene promoter.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; CpG Islands ; DNA Methylation ; Mesenchymal Stromal Cells ; cytology ; Mice ; Octamer Transcription Factor-3 ; metabolism ; Osteoblasts ; cytology ; Promoter Regions, Genetic

OBJECTIVETo isolate, culture and identify of human endometrial stromal stem cells.

METHODSEndometrial tissue was obtained from 10 women undergoing hysterectomy. Purified stromal stem cell suspensions were then obtained by selecting cells with magnetic bead sorting and colony-forming. The surface markers of stromal cell were identified by flow cytometry. Proliferation of stromal stem cell was observed by MTT methods. The osteogenic potential was evaluated by alizarin red staining, the adipogenic potential by oil red O staining. Then the adipogenic and osteogenic specific markers of differentiated cells assayed by RT-PCR method. Expression of cell surface antigen OCT-4 was detected with immunocytochemical staining.

RESULTSEndometrial stem cells were successfully isolated from human endometrial tissue. They were stably proliferated and subcultured in vitro. Most of the passage cells expressed mesenchymal stem cell markers CD90, CD105 but not hemopoietic markers CD34, CD45. The analysis of cell cycle indicated that the percentage of G2-M phase and S phase cells increased. The growth curves of the third passage presented in "S" shape. After cultured in differentiation medium, the cells differentiated toward adipoblasts and osteoblasts as verified by positive staining with Oil Red O and alizarin red staining. Under induction,cells expressed osteogenic and adipogenic marker genes. The immunocytochemical staining of OCT-4 was positive.

CONCLUSIONHuman endometrium contains endometrial stromal stem cells, which present characteristics of mesenchymal stem cells and may be used as seed cells for tissue engineering.

Adult ; Cell Culture Techniques ; Cell Separation ; Cells, Cultured ; Endometrium ; cytology ; Female ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Octamer Transcription Factor-3 ; metabolism ; Stromal Cells ; cytology

With the increasing use of culture-expanded mesenchymal stromal cells (MSCs) for cell therapies, factors that regulate the cellular characteristics of MSCs have been of major interest. Oxygen concentration has been shown to influence the functions of MSCs, as well as other normal and malignant stem cells. However, the underlying mechanisms of hypoxic responses and the precise role of hypoxia-inducible factor-1alpha (Hif-1alpha), the master regulatory protein of hypoxia, in MSCs remain unclear, due to the limited span of Hif-1alpha stabilization and the complex network of hypoxic responses. In this study, to further define the significance of Hif-1alpha in MSC function during their self-renewal and terminal differentiation, we established adult bone marrow (BM)-derived MSCs that are able to sustain high level expression of ubiquitin-resistant Hif-1alpha during such long-term biological processes. Using this model, we show that the stabilization of Hif-1alpha proteins exerts a selective influence on colony-forming mesenchymal progenitors promoting their self-renewal and proliferation, without affecting the proliferation of the MSC mass population. Moreover, Hif-1alpha stabilization in MSCs led to the induction of pluripotent genes (oct-4 and klf-4) and the inhibition of their terminal differentiation into osteogenic and adipogenic lineages. These results provide insights into the previously unrecognized roles of Hif-1alpha proteins in maintaining the primitive state of primary MSCs and on the cellular heterogeneities in hypoxic responses among MSC populations.
*Cell Differentiation ; Cell Proliferation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics/*metabolism ; Kruppel-Like Transcription Factors/genetics/metabolism ; Mesenchymal Stromal Cells/cytology/*metabolism/physiology ; Octamer Transcription Factor-3/genetics/metabolism ; Protein Stability

Cancer stem cells (CSCs) are thought to drive uncontrolled tumor growth, and the existence of CSCs has recently been proven by direct experimental evidence, including tracing cell lineages within a growing tumor. However, CSCs must be analyzed in additional cancer types. Cancer stem cell-like cells (CSCLCs) are a good alternative system for the study of CSCs, which hold great promise for clinical applications. OCT4, NANOG, and SOX2 are three basic transcription factors that are expressed in both CSCLCs and embryonic stem cells (ESCs). These transcription factors play critical roles in maintaining the pluripotence and self-renewal characteristics of CSCLCs and ESCs. In this review, we discuss the aberrant expression, isoforms, and pseudogenes of OCT4, NANOG, and SOX2 in the CSCLC niche, which contribute to the major differences between CSCLCs and ESCs. We also highlight an anticancer therapy that involves killing specific cancer cells directly by repressing the expression of OCT4, NANOG, or SOX2. Importantly, OCT4, NANOG, and SOX2 provide great promise for clinical applications because reducing their expression or blocking the pathways in which they function may inhibit tumor growth and turn-off the cancer "switch." In the future, a clear understanding of transcription factor regulation will be essential for elucidating the roles of OCT4, NANOG, and SOX2 in tumorigenesis, as well as exploring their use for diagnostic and therapeutic purposes.
Animals ; Embryonic Stem Cells ; metabolism ; Homeodomain Proteins ; metabolism ; Humans ; Nanog Homeobox Protein ; Neoplasms ; metabolism ; pathology ; Neoplastic Stem Cells ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; SOXB1 Transcription Factors ; metabolism ; Signal Transduction

PURPOSE: To investigate the association of cancer stem-cell markers [octamer-binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2), and Nanog homebox (NANOG)] expression with clinicopathological properties and overall survival (OS) in operative rectal cancer (RC) patients receiving adjuvant therapy. MATERIALS AND METHODS: 153 patients with primary RC receiving surgery were enrolled. Tumor tissue and paired adjacent normal tissue sample were collected, and OCT4, SOX2, and NANOG expressions were assessed by immunofluorescent staining. The median follow-up duration was 5.2 years, and the last follow-up date was August 2016. RESULTS: Tumor tissue OCT4 (p < 0.001), SOX2 (p=0.003), and NANOG (p < 0.001) expressions were higher than those in adjacent tissue. OCT4 expression was positively correlated with pathological grade (R=0.185, p=0.022), tumor size (R=0.224, p=0.005), and N stage (R=0.170, p=0.036). NANOG expression was positively associated with tumor size (R=0.169, p=0.036). Kaplan-Meier suggested that OCT4+ was associated with worse OS compared with OCT4− (p < 0.001), while no association of SOX2 (p=0.121) and NANOG expressions (p=0.195) with OS was uncovered. Compared with one or no positive marker, at least two positive markers were associated with shorter OS (p < 0.001), while all three positive markers were correlated with worse OS compared with two or less positive markers (p < 0.001). Multivariate Cox's analysis revealed that OCT4+ (p < 0.001) and N stage (p=0.046) were independent factors for shorter OS. CONCLUSION: Tumor tissue OCT4 expression was correlated with poor differentiation, tumor size, and N stage, and it can serve as an independent prognostic biomarker in operative patients with RC receiving adjuvant therapy.
Aged ; Biomarkers, Tumor/metabolism ; Female ; Humans ; Male ; Multivariate Analysis ; Nanog Homeobox Protein/metabolism ; Neoplastic Stem Cells/metabolism ; Octamer Transcription Factor-3/metabolism ; Prognosis ; Rectal Neoplasms/metabolism ; Rectal Neoplasms/pathology ; Rectal Neoplasms/surgery ; SOXB1 Transcription Factors/metabolism ; Survival Analysis

OBJECTIVE: To investigate the expression and significance of hypoxia inducible factor-2α (HIF-2α) and transcription factor OCT-4 in non-small cell lung cancer (NSCLC), and evaluate their roles in the prognosis of NSCLC. METHODS: Tissues from 51 cases of NSCLC were collected and immunohistochemistry (SP method) was used to detect the expression of HIF-2α and OCT-4 proteins. The correlation between the protein expression and the prognosis of NSCLC was analyzed. RESULTS: The positive rates of HIF-2α and OCT-4 expression in the NSCLC were 52.9% and 72.5%, respectively. There was significant relation between the expression of HIF-2α and OCT-4 (r=0.514,P<0.01). High expression of them revealed poor prognosis for NSCLC patients characterized with a bad overall survival(P<0.05). CONCLUSION There is a negative corelation between the expression of HIF-2α and OCT-4 and the prognosis of NSCLC. Combined examination of HIF-2α and OCT-4 expression might be an important biomarker for NSCLC prognosis.
Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Female ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Male ; Octamer Transcription Factor-3 ; metabolism ; Prognosis