OBJECTIVE: To investigate effects of physiological hypoxic conditions on suspension and adherence of embryoid bodies (EBs) during differentiation of human induced pluripotent stem cells (hiPSCs) and explore the underlying mechanisms. METHODS: EBs in suspension culture were divided into normoxic (21% O₂) and hypoxic (5% O₂) groups, and those in adherent culture were divided into normoxic, hypoxic and hypoxia + HIF-1α inhibitor (echinomycin) groups. After characterization of the pluripotency with immunofluorescence assay, the hiPSCs were digested and suspended under normoxic and hypoxic conditions for 5 days, and the formation and morphological changes of the EBs were observed microscopically; the expressions of the markers genes of the 3 germ layers in the EBs were detected. The EBs were then inoculated into petri dishes for further culture in normoxic and hypoxic conditions for another 2 days, after which the adhesion and peripheral expansion rate of the adherent EBs were observed; the changes in the expressions of HIF-1α, β-catenin and VEGFA were detected in response to hypoxic culture and echinomycin treatment. RESULTS: The EBs cultured in normoxic and hypoxic conditions were all capable of differentiation into the 3 germ layers. The EBs cultured in hypoxic conditions showed reduced apoptotic debris around them with earlier appearance of cystic EBs and more uniform sizes as compared with those in normoxic culture. Hypoxic culture induced more adherent EBs than normoxic culture (P < 0.05) with also a greater outgrowth rate of the adherent EBs (P < 0.05). The EBs in hypoxic culture showed significantly up-regulated mRNA expressions of β-catenin and VEGFA (P < 0.05) and protein expressions of HIF-1 α, β-catenin and VEGFA (P < 0.05), and their protein expresisons levels were significantly lowered after treatment with echinomycin (P < 0.05). CONCLUSION Hypoxia can promote the formation and maturation of suspended EBs and enhance their adherence and post-adherent proliferation without affecting their pluripotency for differentiation into all the 3 germ layers. Our results provide preliminary evidence that activation of HIF-1α/β-catenin/VEGFA signaling pathway can enhance the differentiation potential of hiPSCs.
Echinomycin/metabolism* ; Embryoid Bodies/metabolism* ; Humans ; Hypoxia/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; beta Catenin/metabolism*

OBJECTIVETo investigate the expression of cell surface E-cadherin in leukemia cell and the correlation of cell membrane localization of beta-catenin with E-cadherin expression.

METHODSBone marrow samples from 46 patients with acute leukemia and 17 normal donors were analyzed. Cell surface expression of E-cadherin and membrane localization of beta-catenin were labeled by immunofluorescence and analyzed with a laser scanning confocal fluorescence microscope in 14 specimens.

RESULTSCell surface E-cadherin expression level was significantly lower in leukemia cells (with the median fluorescent intensity of 16.78) than in normal hematopoietic progenitors (26.03). Correlation analysis showed that cell membrane localization of beta-catenin was correlated with E-cadherin expression (r = 0.74, P = 0.002). After E-cadherin was induced to express in leukemic cell by 5-Aza-CdR, membranous expression of beta-catenin was elevated while the nuclear expression reduced, indicating that E-cadherin-mediated adhesions could recruit beta-catenin to cell membrane.

CONCLUSIONThe loss of E-cadherin in leukemia cells may result in beta-catenin translocating to the nuclear and transcriptional activation of its target genes.

Cadherins ; metabolism ; Case-Control Studies ; Cell Membrane ; metabolism ; Humans ; Leukemia ; metabolism ; pathology ; beta Catenin ; metabolism

OBJECTIVETo determine the Wnt/beta-catenin signal pathway in different human prostate cancer cell lines and explore its role in epithelial-mesenchymal transition (EMT) in human prostate cancer.

METHODSWe detected the expressions of beta-catenin, t-GSK3beta and p-GSK3beta in several prostate cancer cell lines (LNCaP, C4, C4-2, C4-2B, IF11, IA8, PC-3 and DU145) with different characteristics of epithelial-mesenchymal transition (EMT) by Western blotting.

RESULTSThere were remarkable differences in the expressions of beta-catenin and p-GSK3beta among the cell lines, with beta-catenin and p-GSK3beta highly expressed in LNCaP, C4, C4-2, C4-2B, IF11 and IA8, lowly expressed in PC-3 and DU145, but no difference was observed in the expressions of t-GSK3beta in all the cell lines.

CONCLUSIONThere are differences in the state of the Wnt/beta-catenin signal pathway among the cell lines with different characters of EMT.

Cell Line, Tumor ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo observe the expressions of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines and explore their relationship with the metastasis of human prostate cancer.

METHODSThe expressions and distribution of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines (IF11 and IA8) were detected by Western blot and immunofluorescent staining.

RESULTSThe expression of E-cadherin was high in LNCaP, but absent in IF11 and IA8, while beta-catenin was expressed highly in IF11 and LA8, but lowly in LNCaP. Immunofluorescent staining showed that E-cadherin was mainly in the membrane of LNCaP, while beta-catenin both in the membrane of LNCaP and in the nuclei of IF11 and IA8.

CONCLUSIONE-cadherin and beta-catenin are differently expressed and distributed in prostate cancer cell lines with different characteristics of epithelial-mesenchymal transition (EMT), and the abnormal activation of the beta-catenin signal pathway may be involved in the EMT of prostate cancer cells.

Cadherins ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism

This study was aimed to investigate the expression of beta-catenin in leukemic cell lines and its relationship with pathogenesis of leukemia, semi-quantitative RT-PCR and Western blot were performed to detect the expression of beta-catenin in a panel of 15 human hematopoietic cell lines (U937, KG1a, Jurkat, K562, Namalwa, HEL, HUT78, Raji, Daudi, CEM, LCL-H, HL-60, NB4, J6-1, Ramos). Immunocytochemistry was performed in some of these cell lines to detect the location of beta-catenin. The results showed that the beta-catenin gene was widely expressed in most leukemic cell lines in various degree, the high expression of beta-catenin was found is U937, KG1a, Jurkat, K562 and Namalwa cells, middle expression of beta-catenin was observed in HEL, HUT78, Raji, Daudi and CEM cells, lower expression of beta-catenin was observed in LCL-H, HL-60, NB4, J6-1 and Ramos cells. The expression level of beta-catenin protein was identical to the expression level of beta-catenin mRNA. The expression of beta-catenin could be found in nuclei of all cells mentioned above, but their levels were different between them. Abundant beta-catenin also could be observed in nuclei of some leukemic cells by immunocytochemistry. It is concluded that overexpression of beta-catenin in leukemia cells, as a key mediator of Wnt signaling transduction pathway, indicates that the Wnt signaling transduction pathway may be aberrantly activated in leukemia.
Humans ; Leukemia ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Tumor Cells, Cultured ; beta Catenin ; metabolism

Animals ; Cadherins ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Models, Biological ; Neoplasm Metastasis ; physiopathology ; beta Catenin ; metabolism

Wnt/beta-catenin is the most important and more understanding pathway in Wnt pathways, which is closely related to pathogenesis and development of many solid tumors. Recently, researches discovered that Wnt/beta-catenin signal pathway may be involved in malignant hematopoiesis, and abnormally activated in many hematological-malignancies. This article reviews the newest studies on relationship between Wnt/beta-catenin signal pathway and hematological malignancies (multiple myeloma, chronic myeloid leukemia, chronic lymphocytic leukemia, acute leukemia and so on) in order to reveal the related pathogenesis of hematological malignancies and provide new opinions for target therapy of these diseases.
Hematologic Neoplasms ; metabolism ; Humans ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism

Wnt/β-catenin is an evolutionarily conserved, complex developmental signal pathway that regulates embryogenesis, cell fate, tissue homeostasis, injury repair, and the pathogenesis of human diseases. Mounting evidence demonstrates that Wnt/β-catenin signaling plays a key role in early nephrogenesis. It is relatively silent in normal adult kidneys but reactivated in a wide variety of animal models of nephropathies and in human kidney diseases. Activation of Wnt/β-catenin after acute kidney injury contributes to proper repair and regeneration of damaged renal tubules. However, sustained activation of this signal cascade is closely related to the development and progression of fibrotic chronic kidney disease. In this paper, we systematically review the components and mechanisms of Wnt/β-catenin signaling and its role in kidney repair and fibrosis after injury. A better delineation of the mechanisms of this pathway will provide novel targets and new strategies for designing effective treatment of various kidney diseases.
Animals ; Fibrosis ; Humans ; Kidney/metabolism* ; Renal Insufficiency, Chronic ; Wnt Signaling Pathway ; beta Catenin/metabolism*

Animals ; Mice ; Colitis, Ulcerative/metabolism* ; beta Catenin ; Carcinogenesis ; Probiotics ; Colitis/metabolism*

OBJECTIVETo explore the relationship between the E-cadherin catenin complex and invasiveness of pituitary adenoma.

METHODSThe expression of E-cadherin catenin complex was determined by immunohistochemistry in 78 cases of human pituitary adenomas including invasive adenoma 44 cases, noninvasive adenoma 34 cases and the relativity of their expressions with hormone-producing, pituitary apoplexy and necrosis or cystoid change, tumor diameter were analyzed.

RESULTSThe invasive group had a significantly lower expression of E-cad and alpha-cat than that of noninvasive group (chi-squared = 13.969, P < 0.01). There was no statistical significance for beta-cat expression between the invasive group and noninvasive group (chi-squared = 0.430, P > 0.05). Moreover, the expressions of beta-cad and alpha-cat were significantly lower in macro-adenoma group than that in micro-adenoma group (chi-squared = 5.038, P < 0.05). The expression of E-cad was significantly lower in endocrine inactive group than that in endocrine active group (chi-squared = 4.614, P < 0.05). The expression of beta-cat was significantly lower in the group with apoplexy and necrosis than that in the group without apoplexy and necrosis (chi-squared = 6.701, P < 0.05).

CONCLUSIONSThe reduction of E-cad catenin complex is related to invasiveness and clinical pathological characteristics.

Adolescent ; Adult ; Aged ; Cadherins ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Pituitary Neoplasms ; metabolism ; pathology ; Young Adult ; alpha Catenin ; metabolism ; beta Catenin ; metabolism