Teratocarcinoma-derived Growth Factor 1 Research Articles

Liver receptor homolog 1 (LRH-1) regulates follicle vasculature during ovulation in mice.

It is well-established that liver receptor homolog 1 (LRH-1/NR5A2) regulates the ovarian function and is required for ovulation and luteinization in mice. In the present experiment, we showed that LRH-1 is required to control vascular changes during ovulation, a novel mechanism of action of this orphan nuclear receptor. Liver receptor homolog 1 (LRH-1/NR5A2) is a key regulator of ovarian function, and recently, it has been suggested that it may regulate changes in follicular angiogenesis, an important event during the ovulatory process and luteal development. In the present experiment, the objective was to determine whether conditional depletion of LRH-1 in mice granulosa cells modified vascular changes during the periovulatory period and to explore the possible mechanisms of this modification. We generated mice (22- to 25-day-old) with specific depletion of LRH-1 in granulosa cells by crossing Lrh1 floxed (Lrh1 f/f) mice with mice expressing Cre-recombinase driven by the anti-Müllerian type II receptor (Amhr2-cre; conditional knockout or cKO mice). We showed that preovulatory follicles of LRH-1 cKO mice had a reduced number of endothelial cells in the theca cell layer at 8 h after human chorionic gonadotropin treatment compared with control (CON) mice. Additionally, mRNA and protein expression of leptin receptor (LEPR), a protein that stimulates angiogenesis in a vascular endothelial growth factor-A (VEGFA)-dependent manner, and teratocarcinoma-derived growth factor-1 (TDGF1), which may directly stimulate endothelial cell function, were reduced in LRH-1 cKO mice as compared to CON after the LH surge. These results showed that LRH-1 is necessary for the correct vascular changes that accompany ovulation in mice and that this effect may be regulated through VEGFA-dependent and VEGFA-independent pathways mediated by LEPR and TDGF1.

Teratocarcinoma-derived growth factor-1 (Cripto-1) is overexpressed in epithelial odontogenic lesions displaying more aggressive behaviour.

Cripto-1 also known as teratoma-derived growth factor 1 (TDGF-1) belongs to the EGF-CFC family of growth factor-like molecules. Cripto-1 is involved with embryonic development and not expressed in adult tissue, but some tumours are accompanied by reactivation. The aim of this study was to evaluate the immunohistochemical expression of Cripto-1 in most common odontogenic cysts and tumours. Thirty ameloblastomas, 30 keratocysts, 30 dentigerous cysts and two ameloblastic carcinomas were evaluated using the polymeric immunoperoxidase technique. Immunohistochemical expressions were analysed by the IRS (immunoreactive score). Statistical analyses were performed by the Kruskal-Wallis and Mann-Whitney tests (p ≤ 0.05). Age ranged from 9 to 75years old, with a prevalence of females (n = 49/53.3%). The mandible was the most affected anatomical site (n = 69/75.0%). Cripto-1 immunoexpression was observed in all ameloblastoma, keratocyst and ameloblastic carcinoma cases, although nine dentigerous cyst cases (30%) were negative. Expression scores were higher in ameloblastoma, keratocyst and ameloblastic carcinoma cases (median ranging from 8 to 11) when compared with dentigerous cyst cases (median of 2), with a statistically significant difference (p < 0.001). Cripto-1 is critically important in the progression of several tumours since it is related to significant cell survival and differentiation pathways. The high expression of Cripto-1 in more aggressive odontogenic lesions suggests that this molecule may be involved in the activation of important pathways related to the etiopathogenesis of these lesions.

Teratocarcinoma-Derived Growth Factor-1 (TDGF-1): A Guide Molecule for Monitoring the Treatment Success of Iraqi Patients with Colorectal Carcinoma

This study was carried out in Al Ramadi Teaching hospital from the period March 2013 till August 2013 on sixty two (62) Iraqi patients with colorectal carcinoma. Their ages range from 8–47 years old complain from disturbances in GIT. All patients were visiting the department of surgery in Al-Ramadi teaching hospital. Five ml venous blood was withdrawn from the patients and placed into a plain test tube without anticoagulant to obtain sera. The blood samples were taken from the patients before and after receiving adjuvant chemotherapy and from the healthy individuals (control group 2). The results obtained regarding estimation of TDGF-1 levels in colorectal carcinoma patients revealed that there is a higher significant difference in their concentrations before treatment (3.0 ng/ml) and after chemotherapy treatment (1.2 ng/ml). TDGF-1 level in control group 2 was (0.1 ng/ml). that means, the expression of this TDGF-1 marker in cancerous patients was higher compared with low in normal tissues make it is the target of chemotherapy so evaluation of this marker during receiving chemotherapy give an effective results to choose the best mode of treatment and exclude the non effective ones.

The effect of silencing teratocarcinoma derived growth factor-1 gene on migration of the prostate cancer cell

Objective To study the effect of teratocarcinoma derived growth factor-1 (TDGF-1) siRNA on the ability of migration in prostate cancer cell, and its possible mechanism. Methods The mRNA and protein were extrated from prostate cancer cell line PC3 respectivelytransfected with TDGF-1 siRNA and negative control siRNA, and treatmented with no special. Then wepurified the extrated mRNA and protein. Detecting the TDGF-1 mRNA and protein expression levels by the methods of quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting. The changes of protein expression of EMT markers in PC3 were detected by Western blotting. The cell migration was examined by transwell assay. Results Compared to the negative control (0.988±0.011) and blank group (1.000±0.000), the expression of TDGF-1 mRNA was significantly (P=0.007, P=0.006) decreased in the experimental group (0.250±0.046). The expression of E-cadherin was increased significantly. By contrast, the significantly decreasing expressions of N-cadherin and vimentin were detected. The number of penetrating the membrane was markedly decreased in the experimental group (84.667±7.572) compared to the negative control (168.333±4.041)and blank group (174.333±10.017). Conclusion Down-regulation of TDGF-1 suppressed the migration of PC3 cell, probably related to the EMT. Key words: Teratocarcinoma derived growth factor-1; Prostate cancer; Small interfering RNA; Epithelial-mesenchymal transition; Migration

Bovine lineage specification revealed by single-cell gene expression analysis from zygote to blastocyst.

Preimplantation embryos undergo zygotic genome activation and lineage specification resulting in three distinct cell types in the late blastocyst. The molecular mechanisms underlying this progress are largely unknown in bovines. Here, we sought to analyze an extensive set of regulators at the single-cell level to define the events involved in the development of the bovine blastocyst. Using a quantitative microfluidics approach in single cells, we analyzed mRNA levels of 96 genes known to function in early embryonic development and maintenance of stem cell pluripotency in parallel in 384 individual cells from bovine preimplantation embryos. The developmental transitions can be distinguished by distinctive gene expression profiles and we identified NOTCH1, expressed in early developmental stages, while T-box 3 (TBX3) and fibroblast growth factor receptor 4 (FGFR4), expressed in late developmental stages. Three lineages can be segregated in bovine expanded blastocysts based on the expression patterns of lineage-specific genes such as disabled homolog 2 (DAB2), caudal type homeobox 2 (CDX2), ATPase H+/K+ transporting non-gastric alpha2 subunit (ATP12A), keratin 8 (KRT8), and transcription factor AP-2 alpha (TFAP2A) for trophectoderm; GATA binding protein 6 (GATA6) and goosecoid homeobox (GSC) for primitive endoderm; and Nanog homeobox (NANOG), teratocarcinoma-derived growth factor 1 (TDGF1), and PR/SET domain 14 (PRDM14) for epiblast. Moreover, some lineage-specific genes were coexpressed in blastomeres from the morula. The commitment to trophectoderm and inner cell mass lineages in bovines occurs later than in the mouse, and KRT8 might be an earlier marker for bovine trophectoderm cells. We determined that TDGF1 and PRDM14 might play pivotal roles in the primitive endoderm and epiblast specification of bovine blastocysts. Our results shed light on early cell fate determination in bovine preimplantation embryos and offer theoretical support for deriving bovine embryonic stem cells.

Two independent epigenetic biomarkers predict survival in neuroblastoma.

BackgroundNeuroblastoma (NB) is the most common extracranial pediatric solid tumor with a highly variable clinical course, ranging from spontaneous regression to life-threatening disease. Survival rates for high-risk NB patients remain disappointingly low despite multimodal treatment. Thus, there is an urgent clinical need for additional biomarkers to improve risk stratification, treatment management, and survival rates in children with aggressive NB.ResultsUsing gene promoter methylation analysis in 48 neuroblastoma tumors with microarray technology, we found a strong association between survival and gene promoter hypermethylation (P = 0.036). Hypermethylation of 70 genes significantly differentiated high-risk survivor patients from those who died during follow-up time. Sixteen genes with relevant roles in cancer biology were further validated in an additional cohort of 83 neuroblastoma tumors by bisulfite pyrosequencing. High promoter methylation rates of these genes were found in patients with metastatic tumors (either stage metastatic (M) or metastatic special (MS)), 18 months or older at first diagnosis, MYCN amplification, relapsed, and dead. Notably, the degree of methylation of retinoblastoma 1 (RB1) and teratocarcinoma-derived growth factor 1 (TDGF1) predicts event-free and overall survival independently of the established risk factors. In addition, low RB1 mRNA expression levels associate with poor prognosis suggesting that promoter methylation could contribute to the transcriptional silencing of this gene in NB.ConclusionsWe found a new epigenetic signature predictive for NB patients’ outcome: the methylation status of RB1 and TDGF1 associate with poorer survival. This information is useful to assess prognosis and improve treatment selection.Electronic supplementary materialThe online version of this article (doi:10.1186/s13148-015-0054-8) contains supplementary material, which is available to authorized users.

Evaluation of developmental toxicity using undifferentiated human embryonic stem cells.

An embryonic stem cell test (EST) has been developed to evaluate the embryotoxic potential of chemicals with an in vitro system. In the present study, novel methods to screen toxic chemicals during the developmental process were evaluated using undifferentiated human embryonic stem (hES) cells. By using surface marker antigens (SSEA-4, TRA-1-60 and TRA-1-81), we confirmed undifferentiated conditions of the used hES cells by immunocytochemistry. We assessed the developmental toxicity of embryotoxic chemicals, 5-fluorouracil, indomethacin and non-embryotoxic penicillin G in different concentrations for up to 7 days. While expressions of the surface markers were not significantly affected, the embryotoxic chemicals influenced their response to pluripotent ES cell markers, such as OCT-4, NANOG, endothelin receptor type B (EDNRB), secreted frizzled related protein 2 (SFRP2), teratocarcinoma-derived growth factor 1 (TDGF1), and phosphatase and tensin homolog (PTEN). Most of the pluripotent ES cell markers were down-regulated in a dose-dependent manner after treatment with embryotoxic chemicals. After treatment with 5-fluorouracil, indomethacin and penicillin G, we observed a remarkable convergence in the degree of up-regulation of development, cell cycle and apoptosis-related genes by gene expression profiles using an Affymetrix GeneChips. Taken together, these results suggest that embryotoxic chemicals have cytotoxic effects, and modulate the expression of ES cell markers as well as development-, cell cycle- and apoptosis-related genes that have pivotal roles in undifferentiated hES cells. Therefore, we suggest that hES cells may be useful for testing the toxic effects of chemicals that could impact the embryonic developmental stage.

206 ESTIMATION OF EMBRYO DEVELOPMENTAL TOXICANTS IN HUMAN EMBRYONIC STEM CELL

An embryonic stem cell test (EST) has been developed to evaluate the embryotoxic potential of chemicals with an in vitro system. In the present study, novel methods to screen toxic chemicals during the developmental process were evaluated using undifferentiated human embryonic stem (hES: H9) cells. hES H9 cells were obtained from WiCell Research Institute, Inc., (Madison, WI, USA) and cultured according to the manufacturer's protocol. The cells were cultured on mouse embryonic fibroblast (mEF) cells in hES cell medium composed of DMEM/F12 supplemented with 20% defined fetal bovine serum (FBS), 1% minimal essential medium (MEM) nonessential amino acids, 0.1 mM b-mercaptoethanol, 2 mM L-glutamine, 50 U mL–1 penicillin/streptomycin, and 4 ng mL–1 human recombinant basic fibroblast growth factor. By using surface marker antigens (SSEA-4, TRA-1-60, and TRA-1-81), we confirmed undifferentiated conditions of the used hES cells by immunocytochemistry. We assessed the developmental toxicity of well-known embryotoxic chemicals (cytosine arabinoside, 5-fluorouracil, hydroxyurea, and indomethacin) by expression of pluripotent ES cell markers (OCT-4, NANOG, endothelin receptor type B (EDNRB), secreted frizzled related protein 2(SFRP2), teratocarcinoma-derived growth factor 1 (TDGF1), and phosphatase, and tensin homologue (PTEN)) in different concentrations for up to 7 days. Expression of pluripotent ES cell markers were determined by quantitative real-time PCR. Data are presented as the mean ± standard error of the mean (s.e.m.), and were analysed with a one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test. P-values &lt;0.05 were considered to be statistically significant. Although expressions of the surface markers were not significantly affected, the embryotoxic chemicals influenced their response to pluripotent ES cell markers. Most of the pluripotent ES cell markers were down-regulated in dose-dependent manner following treatment with embryotoxic chemicals. After treatment with 5-fluorouracil, indomethacin, and penicillin G, we observed a remarkable convergence in the degree of up-regulation of development, cell cycle, and apoptosis-related genes by gene expression profiles using Affymetrix GeneChips (Santa Clara, CA, USA). Taken together, these results suggest that embryotoxic chemicals have cytotoxic effects, and modulate the expression of ES cell markers as well as development-, cell cycle-, and apoptosis-related genes that have pivotal roles in undifferentiated hES cells. Therefore, we suggest that hES cells may be useful for testing the toxic effects of chemicals that could affect the embryonic developmental stage.

Abstract A184: Knock-down of Cripto-1 expression by small interfering RNA inhibits growth and migration of oral squamous cell carcinoma cells.

Abstract Background: Cripto-1, also known as teratocarcinoma-derived growth factor-1 (TDGF-1), is a member of the EGF-CFC family. Cripto-1 has been found to be overexpressed in different human carcinomas, including breast, colon, lung, cervix, stomach, pancreas, and nasopharynx, while it was absent or very low in normal adult tissue. In vitro and in vivo studies have shown that Cripto-1 plays an important oncogenic role during carcinogenesis by promoting cell proliferation, migration, invasion, and tumor angiogenesis, as well as inducing the epithelial-to-mesenchymal transition. In our previous study, we first demonstrated that Cripto-1 protein was significantly overexpressed in oral squamous cell carcinoma (OSCC) tissues than in normal oral mucosa and the exogenous recombinant human Cripto-1 protein induced a significant increase in the cell proliferation and migration of OSCC cell lines (SCC-4 and SCC-25). Our data suggested that Cripto-1 may be involved in the tumorigenesis and progression of OSCC. To validate the value of Cripto-1 as a novel molecular target for OSCC treatment, in the present study, we investigated whether the suppression of endogenous Cripto-1 inhibits the growth and migration of OSCC cells. Methods: We screened 13 OSCC cell lines to select cell lines with high Cripto-1 expression by RT-PCR and western blot analysis. Among them, KOSCC-33A and HSC4 cell lines were selected and transfected with anti-Cripto-1 siRNA and negative control siRNA. Cell proliferation was evaluated 24h, 48h, and 72h after transfection, and migration assay was also performed 24h and 48h after transfection, with anti-Cripto-1 siRNA and negative control siRNA. Statistical significance was determined using t-test. Results: The results of RT-PCR analysis revealed that the Cripto-1 mRNA in KOSCC-33A and HSC4 cells were effectively suppressed by anti-Cripto-1 siRNA when compared with cells treated with a negative control siRNA. In the cell proliferation assay, we found that Cripto-1 knock-down induced a significant inhibition of the growth of KOSCC-33A and HSC4 cells (P &amp;lt;0.05). In addition, Cripto-1 knock-down resulted in a significant decrease in the migration of both cell lines (P &amp;lt;0.05). Conclusion: The present study confirms that Cripto-1 plays an important role in the growth and motility of OSCC cells. Our data also suggest that suppression of Cripto-1 expression may be one of the new strategies for the treatment of OSCC. Further studies are in progress to clarify the signaling pathway underlying the oncogenic effects of Cripto-1 in OSCC. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A184. Citation Format: Ji-Hoon Kim, Young-Ah Cho, Ji-Soo Hong, Eun-Jin Choi, Seong-Doo Hong, Jae-Il Lee, Sam-Pyo Hong, Hye-Jung Yoon. Knock-down of Cripto-1 expression by small interfering RNA inhibits growth and migration of oral squamous cell carcinoma cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A184.

Integrating Genetics and Genomics in Primary Aldosteronism

Primary aldosteronism (PA) is the most common form of secondary hypertension, with an estimated prevalence of ≈10% in referred patients and 4% in primary care1,2 but as high as 20% in patients with resistant hypertension.3,4 It is characterized by hypertension with low plasma renin and elevated aldosterone that is often seen with hypokalemia. PA occurs as a result of a dysregulation of the normal mechanisms controlling adrenal aldosterone production. The 2 major causes are aldosterone-producing adenomas (APAs) and bilateral adrenal hyperplasia (BAH), also called idiopathic hyperaldosteronism. The early detection of PA has an enormous impact on clinical outcome and survival, given the major cardiovascular adverse effects of aldosterone excess, which are independent of blood pressure,5,6 and predict outcome after surgical treatment.7,8 Patients with PA have a significantly higher risk of nonfatal myocardial infarction, atrial fibrillation, and stroke compared with age-, sex-, and blood pressure–matched essential hypertensives.5 Furthermore, changes in cardiac structure and function9 and partially reversible renal dysfunction10 have been described, whereas the increased occurrence of metabolic abnormalities in patients with PA is still a matter of debate.11,12 Although the 2008 guidelines for the management of PA have been pivotal for homogenizing screening procedures and treatment among specialized centers,13 there remain a few critical issues related to diagnosis, subtype differentiation, and treatment of nonsurgically correctable forms. Reliable diagnostic and prognostic biomarkers are lacking for more sensitive and specific screening, as well as new therapeutic avenues, because medical and/or surgical treatment of PA leads to normotension in only a minority of patients. This may come from a better understanding of the pathogenic mechanisms of the disease, in particular, identification of the genetic and molecular determinants leading to the development of APA and …

Aldosterone-Producing Adenomas

Primary aldosteronism is the most common form of secondary hypertension and is present in about 8% to 10% of patients with hypertension. In primary aldosteronism, aldosterone secretion is excessive and relatively autonomous of the normal regulatory mechanisms.1,2 It was originally described by Jerome Conn from the University of Michigan3 and is thought to be due to an aldosterone-producing adenoma (APA) causing hypertension and hypokalemic alkalosis. Primary aldosteronism is now recognized to include a spectrum of disorders from unilateral APA to bilateral zona glomerulosa (ZG) hyperplasia, or idiopathic hyperaldosteronism. The diagnosis of all forms of primary aldosteronism is based on the finding of elevated levels of aldosterone and suppressed renin secretion.1 Differentiation between unilateral and bilateral aldosteronism is done by a combination of adrenal imaging (computed tomography or MRI) and bilateral adrenal sampling.1,2 Diagnosis of an APA is attractive because surgical therapy with unilateral adrenalectomy should cure the disease, and mechanistically, APA would suggest a single etiology of a benign neoplastic transformation of cells of the ZG producing aldosterone, regardless of the molecular mechanism. However, the histological characteristics of most adrenal tumors classified as APA are heterogeneous when one considers the surrounding adrenal cortex4 and include varying proportions of 4 different types of cells: clear cells with large vacuolated lipid–laden cytoplasm and central round nuclei similar to zona fasciculata cells; lipid-poor ZG-like cells; compact eosinophilic cells similar to those of the zona reticularis; and cells with cytological features of both ZG and zona fasciculata cells, designated hybrid cells.4 The lipid-laden fasciculata-like cells usually predominate, giving the tumors a characteristic golden yellow color, but some have more glomerulosa-like characteristics.4 However, the histological picture is far more complex, as most patients with APA also have hyperplasia of the rest of the adrenal ZG (40%) or hyperplastic …

Teratocarcinoma-Derived Growth Factor-1 Is Upregulated in Aldosterone-Producing Adenomas and Increases Aldosterone Secretion and Inhibits Apoptosis In Vitro

Aldosterone-producing adenomas (APA) are a frequent cause of secondary hypertension characterized by autonomous hypersecretion of aldosterone. However, the molecular mechanisms involved in adrenal tumorigenesis and deregulated aldosterone secretion are currently unknown. To identify putative functional genes, a transcriptional screening was performed on 8 APA and 3 normal adrenals (NA) using oligonucleotide microarrays. Data were next validated on an expanded set of samples by real-time PCR (APA, n=19; NA, n=10). The epidermal growth factor-like teratocarcinoma-derived growth factor-1 (TDGF-1) was upregulated in APA compared with NA (14.7-fold and 21.4-fold by microarray and real-time PCR, respectively). In vitro studies and Western blot analysis using the NCI H295R adrenocortical cell line showed that TDGF-1 increased Akt phosphorylation on Thr308 and Ser473, consistent with activation of phosphatidylinositol 3-kinase/Akt signaling, and also demonstrated a concomitant inactivation of the Akt substrate glycogen synthesis kinase-3beta via Ser9 phosphorylation. Furthermore, TDGF-1 mediated a 3.8+/-0.4-fold increase in aldosterone secretion (n=4) that was specifically blocked by the phosphatidylinositol 3-kinase inhibitors wortmannin (50 nmol/L) and LY294002 (20 micromol/L). Finally, TDGF-1 protected H295R cells from apoptosis induced by staurosporine, causing a decrease in caspase-3 activity, a reduction in the inactivation of poly(ADP-ribose) polymerase, and an inhibition of DNA fragmentation, detected by the TUNEL reaction and fluorescence microscopy that was blocked by LY294002. Taken together, our data suggest that TDGF-1, which is significantly upregulated in APA and mediates aldosterone hypersecretion and deregulated growth in adrenocortical cells in vitro, may represent a key player in the development and pathophysiology of primary aldosteronism.

TDGF1 is a novel predictive marker for metachronous metastasis of colorectal cancer

Teratocarcinoma-derived growth factor 1 (TDGF1) is a member of the epidermal growth factor-cripto FRL1 cryptic protein family and is involved in the activation of several different signaling pathways during embryonic development and cellular transformation. Previous reports show that TDGF1 regulates the activation of several signaling pathways and controls cellular transformation in embryonic status, whereas its significance in colorectal cancer (CRC) is not yet fully understood. The present study comprised 55 patients who underwent surgery for CRC, as well as two cell lines derived from human CRC. The correlation of gene expression with clinical parameters in patients was assessed. The biological significance of TDGF1 expression was evaluated by knockdown experiments in the cell lines. Seventeen of 55 (30.9%) cases exhibited a higher TDGF1 expression in cancerous regions than in marginal non-cancerous regions. Patients with high TDGF1 expression were statistically susceptible to a recurrence of the disease, and showed poorer disease-free survival than those with low expression. The assessment of TDGF1 knock-down in the 2 cell lines demonstrated that the siRNA inhibition resulted in a statistically significant reduction in cell growth and invasion. In conclusion, the present data strongly suggest the usefulness of TDGF1 as a predictive marker for metachronous metastasis in CRC patients.

Teratocarcinoma-derived growth factor 1 (TDGF1) sequence variants in patients with congenital heart defect

The goal of our study was to identify potential pathogenic mutations in the TDGF1 gene in Chinese people with isolated CHD, particularly those with VSD, and to provide further insight into the etiology of CHD. A total of 500 CHD Chinese patients were investigated for mutations in the TDGF1 gene. Thirteen variants were found among the 500 isolated VSD patients and 250 controls, including one non-synonymous variant identified in patients but not in controls. This work firstly provides human genetic evidence of TDGF1 involved in the pathogenesis of VSD, expanding our knowledge of the causative mutations of congenital heart defects, in particular, the causative mutations of VSD.

Regulation of granulosa and theca cell transcriptomes during ovarian antral follicle development.

Coordinated interactions between ovarian granulosa and theca cells are required for female endocrine function and fertility. To elucidate these interactions the regulation of the granulosa and theca cell transcriptomes during bovine antral follicle development were investigated. Granulosa cells and theca cells were isolated from small (<5 mm), medium (5-10 mm), and large (>10 mm) antral bovine follicles. A microarray analysis of 24,000 bovine genes revealed that granulosa cells and theca cells each had gene sets specific to small, medium and large follicle cells. Transcripts regulated (i.e., minimally changed 1.5-fold) during antral follicle development for the granulosa cells involved 446 genes and for theca cells 248 genes. Only 28 regulated genes were common to both granulosa and theca cells. Regulated genes were functionally categorized with a focus on growth factors and cytokines expressed and regulated by the two cell types. Candidate regulatory growth factor proteins mediating both paracrine and autocrine cell-cell interactions include macrophage inflammatory protein (MIP1 beta), teratocarcinoma-derived growth factor 1 (TDGF1), stromal derived growth factor 1 (SDF1; i.e., CXCL12), growth differentiation factor 8 (GDF8), glia maturation factor gamma (GMFG), osteopontin (SPP1), angiopoietin 4 (ANGPT4), and chemokine ligands (CCL 2, 3, 5, and 8). The current study examined granulosa cell and theca cell regulated genes associated with bovine antral follicle development and identified candidate growth factors potentially involved in the regulation of cell-cell interactions required for ovarian function.

β-Catenin/TCF/LEF regulate expression of the short form human Cripto-1

The human gene Teratocarcinoma-derived growth factor 1 (TDGF1)/Cripto-1/CR-1 which is expressed in a wide variety of human carcinomas is a member of the EGF-cripto FRL1 cryptic (EGF-CFC) gene family. A majority of human colorectal tumors and hepatomas are known to possess a constitutively active canonical Wnt/β-catenin/TCF signaling pathway, also express CR-1. Expression of a short form of CR-1 mRNA in colon carcinoma and hepatoma cell lines suggests that there may be differential regulation of CR-1 expression by the canonical Wnt signaling pathway in colon cancer as well as hepatoma cell lines. The present study demonstrates a direct transcriptional regulation of the short form CR-1 expression by the canonical Wnt signaling pathway through an intronic–exonic enhancer element, containing three tandem TCF/LEF binding sites within the CR-1 gene.

Vascular gene expression and phenotypic correlation during differentiation of human embryonic stem cells

The study of the cascade of events of induction and sequential gene activation that takes place during human embryonic development is hindered by the unavailability of postimplantation embryos at different stages of development. Spontaneous differentiation of human embryonic stem cells (hESCs) can occur by means of the formation of embryoid bodies (EBs), which resemble certain aspects of early embryos to some extent. Embryonic vascular formation, vasculogenesis, is a sequential process that involves complex regulatory cascades. In this study, changes of gene expression along the development of human EBs for 4 weeks were studied by large-scale gene screening. Two main clusters were identified-one of down-regulated genes such as POU5, NANOG, TDGF1/Cripto (TDGF, teratocarcinoma-derived growth factor-1), LIN28, CD24, TERF1 (telomeric repeat binding factor-1), LEFTB (left-right determination, factor B), and a second of up-regulated genes such as TWIST, WNT5A, WT1, AFP, ALB, NCAM1. Focusing on the vascular system development, genes known to be involved in vasculogenesis and angiogenesis were explored. Up-regulated genes include vasculogenic growth factors such as VEGFA, VEGFC, FIGF (VEGFD), ANG1, ANG2, TGFbeta3, and PDGFB, as well as the related receptors FLT1, FLT4, PDGFRB, TGFbetaR2, and TGFbetaR3, other markers such as CD34, VCAM1, PECAM1, VE-CAD, and transcription factors TAL1, GATA2, and GATA3. The reproducibility of the array data was verified independently and illustrated that many genes known to be involved in vascular development are activated during the differentiation of hESCs in culture. Hence, the analysis of the vascular system can be extended to other differentiation pathways, allocating human EBs as an in vitro model to study early human development.

A Truncated Form of Teratocarcinoma-Derived Growth Factor-1 (Cripto-1) mRNA Expressed in Human Colon Carcinoma Cell Lines and Tumors

The human teratocarcinoma-derived growth factor (TDGF)-1 gene encodes a 188-amino acid protein, cripto-1. The TDGF-1 gene is overexpressed in the majority of human primary colorectal carcinomas and hepatic metastases, in breast carcinomas and in testicular nonseminoma germ cell embryonal carcinomas. In the human embryonal carcinoma cell line NTERA-2 clone D1, a 2-kb TDGF-1 mRNA transcript is expressed. The present study shows that a 1.7-kb mRNA transcript lacking the first two exons of the TDGF-1 gene is expressed in the human colon carcinoma cell line GEO. This shorter mRNA is the only TDGF-1 transcript that is present in the majority of primary human colorectal carcinomas and hepatic metastases and in adult human tissues such as the pancreas, heart, stomach, mammary gland, skeletal muscle, liver and placenta. In contrast, in the kidney, brain, testis, ovary and spleen, the longer 2-kb TDGF-1 mRNA transcript is expressed. The putative shorter protein starts at a CUG codon 129 nucleotides downstream of the starting AUG codon of the longer protein. These data indicate the potential for differential transcriptional regulation of the TDGF-1 gene in different normal and tumor tissues.

Cripto: a novel epidermal growth factor (EGF)-related peptide in mammary gland development and neoplasia.

Growth and morphogenesis in the mammary gland depend on locally derived growth factors such as those in the epidermal growth factor (EGF) superfamily. Cripto-1 (CR-1, human; Cr-1, mouse)--also known as teratocarcinoma-derived growth factor-1--is a novel EGF-related protein that induces branching morphogenesis in mammary epithelial cells both in vitro and in vivo and inhibits the expression of various milk proteins. In the mouse, Cr-1 is expressed in the growing terminal end buds in the virgin mouse mammary gland and expression increases during pregnancy and lactation. Cr-1/CR-1 is overexpressed in mouse and human mammary tumors and inappropriate overexpression of Cr-1 in mouse mammary epithelial cells can lead to the clonal expansion of ductal hyperplasias. Taken together, this evidence suggests that Cr-1/CR-1 performs a role in normal mammary gland development and that it might contribute to the early stages of mouse mammary tumorigenesis and the pathobiology of human breast cancer.

Assignment of human teratocarcinoma derived growth factor (TDGF) sequences to chromosomes 2q37, 3q22, 6p25 and 19q13.1

The human teratocarcinoma derived growth factor 1 (TDGF1) gene maps on chromosome (Chr) 3p21.3. One pseudogene (TDGF3) maps on Chr Xq21→q22. We now report the nucleotide sequence and chromosome location of three additional TDGF pseudogenes. The three new sequences (TDGF2, TDGF4 and TDGF5) are truncated at the 5′ end and have accumulated several point mutations, deletions and insertions. TDGF2, TDGF4 and TDGF6 map on Chrs 2q37, 6p25 and 3q22, respectively. Finally, Southern blot analysis of DNA from normal individuals shows a highly variable restriction pattern of the TDGF sequences.