CDX2 Gene Expression Research Articles

CDX1 Gene Expression in Recurrent Spontaneous Abortion under Extracellular Regulatory Protein Kinase1/2 Signaling Pathway

CDX1 Gene Expression in Recurrent Spontaneous Abortion under Extracellular Regulatory Protein Kinase1/2 Signaling Pathway

CDX2 protein distribution in the gastric mucosa in chronic gastritis by a semi-quantitative index and its reproducibility

Introduction The Cdx2 gene provides an intestinal differentiation of epithelial cells and plays an oncosupressive role. An indirect method of the Cdx2 gene expression assessment is the immunohistochemical study of its product, the CDX2 protein. Therefore, the common approach to the immunohistochemical study of the CDX2 protein hasn’t been developed yet. A semi-quantitative CDX2 index based on the percentage of CDX2-positive cells in the tissue specimen, the staining intensity and an expression pattern has been proposed.The purpose of the study was to assess the reproducibility of the semi-quantitative CDX2 index calculation in chronic atrophic gastritis stages I-IV.Materials and methods 20 chronic atrophic gastritis cases (5 cases for each stage of the gastritis according to the Operative link for gastritis assessment system) were taken according to the Maastricht V protocol and examined by the immunohistochemical method (CDX2, clone EPR2764Y, ready to use). The reproducibility of the CDX2 semi-quantitative index was assessed by five pathologists. An agreement between observed raters was measured by the kappa statistics.Results The Cohen’s κ value is 0,8 for unweighted κ and 0,97 for weighted κ (extremely high level of agreement) for the semi-quantitative CDX2 index calculation.Discussion The least reproducible parameter used for the semi-quantitative CDX2 index calculation the percentage of CDX2-positive cells because of the subjective assessment. The Cohen’s weighted κ value was higher compared to the unweighted κ because of the close yet not similar CDX2 semi-quantitative index values calculated by pathologists.Conclusion The semi-quantitative CDX2 index can be used to rank CDX2 expression and has a high level of reproducibility.

Targeted silencing of CDX2 gene with siRNA enhanced vincristine-induced inhibition of proliferation and induction of apoptosis of leukemia K562 cells

Purpose: To determine the effect of targeted silencing of CDX2 with siRNA on vincristine-induced inhibition of proliferation and apoptosis induction in leukemia K562 cells.
 Methods: K562 cells were divided into untreated group, vincristine group, blank group, and CDX2-siRNA group. The expression of CDX2 gene after CDX2-siRNA transfection was determined by reverse-transcription-polymerase chain reaction (RT-PCR) and immunoblotting, while MTT and flow cytometric procedures were used to evaluate the effect of vincristine on proliferation and apoptotic changes in K562 cells.
 Results: Protein expression of CDX2 did not change significantly in non-treated cells, vincristine group and blank group, while the expression of CDX2 protein in cells of CDX2-siRNA group was decreased significantly (p < 0.05) while MTT assay results showed that the absorbance of CDX2-siRNA cells was significantly lower than those of the other three groups at 12, 24 and 48 h after CDX2 gene silencing. Flow cytometry showed markedly higher percentage apoptosis of CDX2-siRNA group than in other groups at 12, 24 and 48 h after CDX2 gene silencing. There was no difference in apoptosis level between vincristine group and blank group, but apoptosis was higher in these groups than in untreated group (p < 0.05).
 Conclusion: Silencing CDX2 gene via targeting with siRNA enhances vincristine-induced suppression of growth and apoptotic changes in leukemia K562 cells, thereby enhancing the anti-tumor effect of vincristine.

Cryoprotectant role of exopolysaccharide ID1 in the vitrification/in-straw warming of in vitro-produced bovine embryos.

The cold-adapted bacterium Pseudomonas sp. ID1 produces the extracellular exopolysaccharide ID1 (EPS ID1) with cryoprotective activity. This study was designed to optimize the vitrification/in-straw warming protocol of in vitro-produced (IVP) blastocysts by adding EPS ID1 to the vitrification media. Day 7-expanded blastocysts were vitrified/warmed using the VitTrans device after the addition of 0 or 100 μg/mL EPS ID1 to the vitrification media. Blastocysts vitrified by the Cryotop method and fresh non-vitrified blastocysts served as controls. Outcomes were assessed in the warmed embryos in terms of survival rates and mRNA relative abundances of BAX, BCL2, GPX1, and CDX2 genes. No differences in survival rates were observed at 3h post-warming between vitrification treatments. At 24 h post-warming, the addition of EPS prior to vitrification with the VitTrans device produced similar survival rates to Cryotop-vitrified embryos and similar hatching rates to fresh non-vitrified or Cryotop-vitrified embryos. No differences emerged in BCL2 gene expression. Lower BAX (p < .05) and higher GPX1 (p < .05) and CDX2 (p < .1) gene expression were observed in expanded and/or hatched blastocysts derived from VitTrans-EPS-vitrified embryos when compared to those from the non-supplemented group. In conclusion, addition of EPS not only promoted blastocyst survival and hatching after VitTrans vitrification/warming but also modified the expression of genes associated with better embryo quality.

The effects of embryo splitting on Cdx2, Sox2, Oct4, and Nanog gene expression in mouse blastocysts.

Embryo splitting is utilized in reproduction biotechnology. The blastomeres resulting from the splitting of an embryo in two-, four- or eight-cell stages can develop into separate embryos that are genetically similar to the other blastomeres. The present work studied the effects of splitting on embryo pluripotent gene expression (Cdx2, Sox2, Oct4, and Nanog) in mice. Two-cell embryos were isolated from stimulated mice. The embryos were grouped into "split" and "non-split" groups. The zona pellucida was removed from the split group and the blastomeres were distributed before being co-cultured with mouse embryo fibroblasts to the blastocyst stage. Normal (non-split) blastocysts were co-cultured in the same way. The 3.5-day-old blastomeres were collected as the control group. For molecular evaluation, real-time PCR was conducted to analyze changes in Cdx2, Sox2, Oct4, and Nanog gene expression. Moreover, the blastocyst formation rate, overall blastocyst rate, and the number of newborns were statistically analyzed. The findings showed that embryo splitting increased blastocyst formation, overall blastocysts, developmental potential embryos, and the number of infants. Furthermore, the split and non-split (control) groups showed equal expression of pluripotent genes (Cdx2, Sox2, Oct4, and Nanog) in the molecular analysis. It can be concluded that the growth and developmental potency of sister blastocysts derived from split two-cell stage mouse embryos are the same as those of normal blastocysts. So, there are no significant differences in gene expression between the split and non-split groups.

385 MIR-24-3P REGULATES CDX2 DURING THE PROCESS OF INTESTINALIZATION OF THE CARDIAC-TYPE EPITHELIUM IN A HUMAN MODEL OF BARRETT’S ESOPHAGUS

Abstract Cardiac-type epithelium has been proposed as an intermediate stage between normal squamous epithelium and intestinal metaplasia in the development of Barrett’s esophagus. Deregulation of certain miRNAs and their effects on CDX2 expression might contribute to the intestinalization process of cardiac-type epithelium. The aim of this study was to identify miRNAs differentially expressed between CDX2 positive and negative glands of Barrett’s esophagus and to examine the function of specific miRNAs on the regulation of CDX2. Methods miRNA expression profiling using OpenArrayTM analysis in microdissected cardiac-type glands with and without fully CDX2 expression was performed in biopsies from patients who developed cardiac-type epithelium in the remnant esophagus after esophagectomy. Data were validated using real-time PCR in esophageal adenocarcinoma cell lines and in situ and real-time PCR miRNA/CDX2/MUC2 co-expression analysis in cardiac-type mucosa samples. The effect of miR-24-3p precursor transfection on CDX2 expression was assessed in the esophageal adenocarcinoma cell lines FLO-1 and KYAE-1. Results CDX2 positive glands were characterized by an unique miRNA profile with a significant downregulation of miR-24-3p, miR-520e-3p, miR-548a-1, miR-597-5p, miR-133a-3p, miR-30a-5p, miR-638, miR-625-3p, miR-1255b-1, miR-1260a and upregulation of miR-590 (Figure 1A). miRNA-24-3p was identified as potential regulator of CDX2 gene expression in three bioinformatics algorithms, and this was confirmed in esophageal adenocarcinoma cell lines (Figure 1C). Furthermore, miR-24-3p expression negatively correlates with CDX2 in cardiac-type mucosa samples with different stages of mucosal intestinalization (Figure 1B). Conclusion These results imply that miRNA-24-3p directly targets CDX2, and downregulation of miRNA-24-3p is associated with the acquisition of an intestinal phenotype in cardiac-type epithelium.

MiR-24-3p regulates CDX2 during intestinalization of cardiac-type epithelium in a human model of Barrett's esophagus.

Cardiac-type epithelium has been proposed as the precursor of intestinal metaplasia in the development of Barrett's esophagus. Dysregulation of microRNAs (miRNAs) and their effects on CDX2 expression may contribute to intestinalization of cardiac-type epithelium. The aim of this study was to examine the possible effect of specific miRNAs on the regulation of CDX2 in a human model of Barrett's esophagus. Microdissection of cardiac-type glands was performed in biopsy samples from patients who underwent esophagectomy and developed cardiac-type epithelium in the remnant esophagus. OpenArray™ analysis was used to compare the miRNAs profiling of cardiac-type glands with negative or fully positive CDX2 expression. CDX2 was validated as a miR-24 messenger RNA target by the study of CDX2 expression upon transfection of miRNA mimics and inhibitors in esophageal adenocarcinoma cell lines. The CDX2/miR-24 regulation was finally validated by in situ miRNA/CDX2/MUC2 co-expression analysis in cardiac-type mucosa samples of Barrett's esophagus. CDX2 positive glands were characterized by a unique miRNA profile with a significant downregulation of miR-24-3p, miR-30a-5p, miR-133a-3p, miR-520e-3p, miR-548a-1, miR-597-5p, miR-625-3p, miR-638, miR-1255b-1, and miR-1260a, as well as upregulation of miR-590-5p. miRNA-24-3p was identified as potential regulator of CDX2 gene expression in three databases and confirmed in esophageal adenocarcinoma cell lines. Furthermore, miR-24-3p expression showed a negative correlation with the expression of CDX2 in cardiac-type mucosa samples with different stages of mucosal intestinalization. These results showed that miRNA-24-3p regulates CDX2 expression, and the downregulation of miRNA-24-3p was associated with the acquisition of the intestinal phenotype in esophageal cardiac-type epithelium.

15 Blastocysts altered CDX2 and SOX2 gene expression and pregnancy failure after embryo transfer in yak heterospecific somatic cell nuclear transfer

Heterospecific cloning is a tool for the genetic rescue of endangered animals. Our objective was to evaluate the effects of heterospecific yak (Bos grunniens) cloned embryo aggregation on the expression levels of NANOG, OCT4, CDX2, and SOX2 genes, and to compare with IVF, parthenogenetic zona-free (P-ZF), and homospecific bovine cloned embryos (BB1x). Oocytes were recovered from the ovaries of slaughtered cows and invitro matured for 22h. The zona pellucida was removed by protease treatment and then mature oocytes were enucleated by micromanipulation. Enucleated oocytes were placed in phytohemagglutinin to induce adherence with a somatic donor cell followed by electrofusion (with two 30-µs pulses of 1.2 kV/cm, 0.1s apart). Two hours after fusion, reconstructed embryos were activated using ionomycin followed by 6-(dimethylamino)purine (6-DMAP) treatment for 3h and cultured in synthetic oviductal fluid (SOF) medium for 7 days. The experimental groups were IVF, P-ZF, BB1x, heterospecific yak-bovine cloned embryos (1 embryo per microwell, YB1x), and heterospecific yak-bovine cloned embryos aggregated (2 embryos per microwell, YB2x). In all experimental groups, cleavage and blastocyst rates were assessed 7 days after activation. In addition, 5 blastocysts were pooled for each biological replicate, and pluripotency-specific genes (NANOG, SOX2, CDX2, and OCT4) were analysed by quantitative PCR. Data were analysed by the ΔΔCT method using the geometric mean of ACTB (actin) and GAPDH as internal standard followed by one-way ANOVA. Cleavages rates were significantly lower in the YB1x group compared with the other groups. Moreover, blastocyst rates in YB2x (31.34%, n=67) were significantly higher than in YB1x (13.86%, n=101) and BB1x (13.33%, n=45) groups, but there were no significant differences compared with the IVF (43.82%, n=89) and P-ZF (25%, n=68) groups. In contrast, although no significant differences were observed among groups in the expression of NANOG and OCT4 genes, the expression of CDX2 was lower in YB2x and YB1x blastocysts compared with the BB1X, P-ZF, and IVF (control) groups. In addition, a decrease in SOX2 gene expression was observed in the YB2x and YB1x blastocysts compared with the BB1X group. Blastocysts from YB1x (n=5) and YB2x (n=18) groups were transferred to recipient cows (n=23) on Day 7. Forty days after embryo transfer, presence of uterine fluid was detected by ultrasound in 3 recipient cows (from YB2x), suggesting embryo loss. In concordance with our previous reports, yak heterospecific SCNT blastocysts showed underexpression of CDX2 and SOX2 compared with the overexpression observed for these genes in bovine homospecific SCNT blastocysts. Thus, yak heterospecific SCNT blastocysts may have compromised developmental competence associated with altered expression of CDX2 and SOX2 that cannot be rescued by the aggregation of 2 reconstructed embryos.

Expression of Hsa-miR-9 in Children with Acute Lymphoblastic Leukemia and Its Relationship with CDX2 Gene

To investigate the expression of Hsa-miR-9 in children with acute lymphoblastic leukemia (ALL) and its relationship with CDX2 gene. The clinical data of 130 patients with ALL were collected from nearly five years in our hospital, and in the same period 60 healthy children in the same age were selected as control group. Real-time fluorescent quantitative PCR (qRT-PCR) was applied to examine the expression of Hsa-miR-9 and CDX2 gene of the two groups, the relationship between Hsa-miR-9 expression and ALL patients' clinical characteristics, survival time, CDX2 expression were analyzed. The expression level of Hsa-miR-9 in ALL children was significantly lower than that in the healthy control group (P<0.001). The expression of Hsa-miR-9 in ALL children related with serum WBC, infiltrating lymph nodes, splenomegaly, and risk grade (P<0.05). The median survival time of ALL children with high expression of Hsa-miR-9 was significantly longer than that of the ALL children with low expression (P<0.001). Hsa-miR-9 expression significantly correlated with CDX2 gene expression in ALL children at different treatment stages (P<0.05). The expression of Hsa-miR-9 decreases in children with ALL, and ALL children with high expression of Hsa-miR-9 have a longer overall survival time. The expression of hsa-miR-9 in ALL children closely related with CDX2 gene.

DMSO supplementation during in vitro maturation of bovine oocytes improves blastocyst rate and quality

The molecule Dimethyl sulfoxide is widely used as drug solvent. However, its antioxidant property was poorly explored. In this study, we evaluated the effect of DMSO supplementation during oocyte in vitro maturation (IVM) on embryo development and quality. Bovine oocytes were matured with different DMSO concentrations (0, 0.1, 0.25, 0.5, 0.75, 1 and 10% v:v) followed by in vitro fertilization. Subsequently, quality indicators such as gene expression of SOX2, OCT4, CDX2, SOD1, oocyte and embryo redox status and DNA damage were evaluated. Polar body extrusion and blastocyst rates increased with 0.5% v:v DMSO. Moreover, first polar body extrusion and blastocyst rates did not increase with 1%, and 10% of DMSO reduced polar body extrusion and did not produce blastocyst. Optimal concentration of DMSO for the use on the maturation was estimated at around 0.45% v:v. Supplementation with 0.5% v:v DMSO has not affected mRNA abundance of genes key in blastocyst, however 0.75% increased gene expression of OCT4 and SOX2. Oocytes matured with 0.5% v:v DMSO and blastocyst from DMSO group showed reduced lipid peroxidation respect control. Total Glutathione concentrations increased in blastocyst stage in DMSO group. DMSO increased the total cell number of blastocysts but not TUNEL positive cells. In conclusion, our results suggest that low DMSO concentrations used during bovine oocytes in vitro maturation increases the maturation, as well as the blastocyst rate and its quality, without demonstrating deleterious effect on embryo cells.

Development of Physiologically Responsive Human iPSC-Derived Intestinal Epithelium to Study Barrier Dysfunction in IBD.

In inflammatory bowel disease (IBD), the intestinal epithelium is characterized by increased permeability both in active disease and remission states. The genetic underpinnings of this increased intestinal permeability are largely unstudied, in part due to a lack of appropriate modelling systems. Our aim is to develop an in vitro model of intestinal permeability using induced pluripotent stem cell (iPSC)-derived human intestinal organoids (HIOs) and human colonic organoids (HCOs) to study barrier dysfunction. iPSCs were generated from healthy controls, adult onset IBD, and very early onset IBD (VEO-IBD) patients and differentiated into HIOs and HCOs. EpCAM+ selected cells were seeded onto Transwell inserts and barrier integrity studies were carried out in the presence or absence of pro-inflammatory cytokines TNFα and IFNγ. Quantitative real-time PCR (qRT-PCR), transmission electron microscopy (TEM), and immunofluorescence were used to determine altered tight and adherens junction protein expression or localization. Differentiation to HCO indicated an increased gene expression of CDX2, CD147, and CA2, and increased basal transepithelial electrical resistance compared to HIO. Permeability studies were carried out in HIO- and HCO-derived epithelium, and permeability of FD4 was significantly increased when exposed to TNFα and IFNγ. TEM and immunofluorescence imaging indicated a mislocalization of E-cadherin and ZO-1 in TNFα and IFNγ challenged organoids with a corresponding decrease in mRNA expression. Comparisons between HIO- and HCO-epithelium show a difference in gene expression, electrophysiology, and morphology: both are responsive to TNFα and IFNγ stimulation resulting in enhanced permeability, and changes in tight and adherens junction architecture. This data indicate that iPSC-derived HIOs and HCOs constitute an appropriate physiologically responsive model to study barrier dysfunction and the role of the epithelium in IBD and VEO-IBD.

24 Asynchronic tetraploid complementation and embryo quality in domestic cat and Leopardus geoffroyi hybrid embryos

Fusion of 2-cell embryos generates tetraploid (4n) blastomeres with an increased commitment to trophectoderm. Complementation of embryos from endangered species with 4n blastomeres derived from a phylogenetically related domestic species could improve healthy pregnancy establishment after embryo transfer in domestic females. However, generation, development, and quality of tetraploid complemented embryos in felids remain unstudied. Therefore, our objectives were (1) to evaluate tetraploidy of 2-cell fused embryos; (2) to analyse the blastocyst cell number, distribution after synchronic (S) or asynchronic (AS) complementation, OCT4+ cells, DNA-fragmentation levels and CDX2 gene expression of IVF complemented embryos; and (3) to evaluate the developmental rates of tetraploid complemented Felis catus-Leopardus geoffroyi hybrid embryos. After ovariectomy, Felis catus oocytes were IVM and subjected to IVF. For Experiment 1 (n=66), 2-cell embryos (2n) were exposed to two 30-ms DC pulses at 8 kV cm−1 electric field in fusion media. Fused (4n) and nonfused embryos were cultured invitro in 50-μL drops of modified Tyrode's medium. Karyotype analysis was performed at Day 4. For Experiment 2 (n=24), zona-free IVF embryos were aggregated S (4-cell-2n/4-cell-2n) or AS (4-cell-2n/2-cell-2n and 4-cell-2n/1-cell-4n). For Experiment 3 (n=36), blastocysts generated by AS complementation (4-cell-2n/2-cell-2n and 4-cell-2n/1-cell-4n) were either fixed with 4% paraformaldehyde for immunofluorescence and terminal deoxynucleotidyl transferase dUTP nick end labeling assay or saved in RNA-Later for RT-qPCR analysis. For this experiment, nonaggregated 2n and 4n blastocysts were used as a control. For Experiment 4 (n=60), IVM oocytes were co-incubated with Leopardus geoffroyi and Felis catus (control) spermatozoa and then 4-cell-2n heterologous embryos were complemented with 1-cell-4n homologous IVF embryos. Data were analysed by Fisher's exact test. Our results showed that 67% of the 2-cell fused embryos were 4n. Moreover, 82% of nonfused embryos were aneuploids compared with 78% of 2n embryos in the control group. The AS complemented blastocysts (4-cell-2n/1-cell-4n and 4-cell-2n/2-cell-2n) had significantly higher cell number compared with S complemented (4-cell-2n/4-cell-2n) or noncomplemented embryos. The AS complementation also increased the number of OCT4+ cells independently of the ploidy of the embryos. Interestingly, AS tetraploid complemented embryos had significantly lower number of cells with fragmented DNA. No differences were found in CDX2 expression among complemented embryos; however, noncomplemented 2n blastocysts showed a significantly lower expression compared with the others group. Finally, we observed that AS complementation of 2n hybrid embryos with 4n homologous embryos reached similar blastocyst rates, 70 and 88%, respectively. Our findings support the use of 2-cell fused embryos to generate 4n blastomeres and demonstrated that tetraploid complementation generates good quality embryos, providing evidences that encourage the use of this technology to improve the developmental competence of interspecific embryos after transfer.

P160 miR-24-3p REGULATES CDX2 DURING THE PROCESS OF INTESTINALIZATION OF THE CARDIAC-TYPE EPITHELIUM IN A HUMAN MODEL OF BARRETT´S ESOPHAGUS

Abstract Aim The aim of this study was to identify miRNAs differentially expressed between CDX2 positive and negative glands of Barrett’s esophagus and, based on this, to examine the function of specific miRNAs on the regulation of CDX2. Background &amp; Methods Cardiac-type epithelium has been proposed as an intermediate stage between normal squamous epithelium and intestinal metaplasia (IM) in the development of Barrett´s esophagus. Deregulation of certain miRNAs and their effects on CDX2 expression might contribute to the intestinalization process of cardiac-type epithelium. miRNA expression profiling using OpenArray technology in microdissected cardiac-type glands with and without fully CDX2 expression was performed in biopsies from patients who developed cardiac-type epithelium in the remnant esophagus after esophagectomy. Data were validated using real-time PCR in esophageal adenocarcinoma cell lines and in situ miRNA/CDX2 co-expression analysis in cardiac-type mucosa samples. The effect of miR-24-3p precursor transfection on CDX2 expression was assessed in the esophageal adenocarcinoma cell lines FLO-1 and KYAE-1. Results CDX2 positive glands were characterized by an unique miRNA profile with a significant downregulation of miR-24-3p, miR-520e-3p, miR-548a-1, miR-597-5p, miR-133a-3p, miR-30a-5p, miR-638, miR-625-3p, miR-1255b-1, miR-1260a and upregulation of miR-590 (Figure A). miRNA-24-3p was identified as potential regulator of CDX2 gene expression in three bioinformatics algorithms, and this was confirmed in esophageal adenocarcinoma cell lines (Figure C). Furthermore, miR-24-3p expression negatively correlates with CDX2 in cardiac-type mucosa samples with different stages of mucosal intestinalization (Figure B). Conclusion These results imply that miRNA-24-3p directly targets CDX2, and downregulation of miRNA-24-3p is associated with the acquisition of an intestinal phenotype in cardiac-type epithelium. Figure: (A) OpenArray results showing downregulation of 10 miRNAs in CDX2 positive glands and upregulation of one miRNA (miR-590). (B) miR-24-3p in situ hybridization: intestinal metaplastic glands were characterized by a positive staining for CDX2 (red) and faint to negative miR-24-39. Adjacent non-intestinalized CDX2-negative glands showed a moderate miR-24-3p expression. (C) Transfection of KYAE-1 cells with miR-24-3p and miR-24-39 inhibitor. A significant CDX2 decrease was observed.

Immunohistochemical expression of CDX2 gene in colorectal carcinoma

Background and objective: Colorectal cancer is a multi-factorial disease process, with etiology encompassing genetic factors, environmental exposures, and inflammatory conditions of the digestive tract. This study aimed to investigate the immunohistochemical expression of CDX2 as well as clinical and histopathological parameters of patients with colorectal cancer in Erbil, Kurdistan. Methods: A retrospective study including about 100 colorectal cancer cases in Erbil city, Kurdistan was conducted from January 2015 to January 2017. Tumor type, site, size, histological grade, lymph node status, and pathological stage with CDX2 expression were investigated. Results: CDX2 was expressed in 86 out of 100 (86%) patients. It was more expressed (focal or diffuse staining) in nonmucinous carcinoma more than signet ring and mucinous carcinoma. Loss of CDX2 expression in colorectal cancer is associated with a high tumor grade and stage. Significant associations between CDX2 expression with both tumor type and tumor grade were observed. Although CDX2 expression was found to be reduced in proximal location (right colon) and higher stage, however, no significant associations between CDX2 expressions in colorectal cancer were detected with tumor site, nodal status, and tumor stage. Conclusion: Reduced and loss CDX2 expression in colorectal cancer associated with high tumor grades as well as with the mucinous and signet ring cell carcinomas may reflect aggressive tumor behavior probably because of the CDX2 tumor-inhibitory properties. However, the full extent of CDX2’s antitumor effects has yet to be elucidated.

CDX1/2 and KLF5 Expression and Epigenetic Modulation of Sonic Hedgehog Signaling in Gastric Adenocarcinoma.

Gastric cancer is among the commonplace causes of cancer death worldwide. Sonic hedgehog (Shh) signaling is an important pathway which may be dysregulated in many cancers.CDX1/2, and KLF5are key transcription factors involved in Shh pathway and cancer stem cells. The aim of this study was to investigate the expression and epigenetic alterations of these genes in gastric cancer patients. DNA methylation's modifications of CDX1, KLF5 and CDX2 genes alongside with the expressions of these genes in gastric cancer tissues and their non-tumoral counterparts (margin tissues) were analyzed using methylation specific sequencing, and Real time PCR Taq man assays, respectively. The expression of CDX1 (P = 0.002) and KLF5 (P = 0.010) were decreased significantly, but it was considerably increased for CDX2 (P = 0.001). Relatively, the results for the regulatory region methylation status of each CpG site had shown a notable fluctuation in these genes with no significant difference in most places. The creation of metastatic lymph nodes in patients was significantly associated with increased expression of CDX2 gene. The modifications of these genes expression can be considered as a cancer biomarker in future studies. Methylation of the investigated genes is not the main mechanism of gastric cancer development.

Methylation of CDX2 gene promoter in the prediction of treatment efficacy in colorectal cancer.

The aim of the present study was to examine the diagnosis of methylation of CDX2 gene promoter in colorectal cancer (CRC) and assessed its value in the prediction of treatment efficacy. Sixty patients who were diagnosed as CRCs for the first time, 60 patients with hyperplastic polyps (HPs) and adenomas, and 60 patients with inflammatory lesions or healthy patients (control group) were included in the present study. The methylation levels of CDX2 gene promoter were detected by methylation-specific polymerase chain reaction (MSP), and the expression levels of CDX2 mRNA were detected by fluorescence quantitative PCR. Treatment options, such as surgery, radiotherapy and chemotherapy, were chosen on the basis of TNM staging of CRC patients. The tumor-free survival, relapse rate and mortality were also recorded. The methylation rate was 71.67% (43/60) and significantly higher in the CRC group as compared to the HP/adenoma and control groups, P<0.05. Moreover, they showed further increase with higher degree of TNM staging. The expression levels of CDX2 mRNA was significantly lower in the CRC group in comparison to HP/adenoma and control groups, P<0.05, and showed a further decrease with a higher degree of TNM staging. The tumor-free survival was shorter, and the relapse rate and mortality were higher in patients with positive methylation in the CRC group, P<0.05. Multivariate logistic regression analysis demonstrated that TNM staging and positive methylation were independent risk factors of mortality. In conclusion, higher methylation degree of CDX2 gene promoter resulted in decreased expression of CDX2 gene, and was closely associated with TNM staging and prognosis. TNM staging and positive methylation were independent risk factors of mortality for CRC patients.

HIGH CDX2 GENE EXPRESSION PREDICTS INFERIOR PROGNOSIS IN ACUTE MYELOID LEUKEMIA PATIENTS WITH NORMAL CYTOGENETICES.

20Dec 2016 HIGH CDX2 GENE EXPRESSION PREDICTS INFERIOR PROGNOSIS IN ACUTE MYELOID LEUKEMIA PATIENTS WITH NORMAL CYTOGENETICES. Shereen Mohamed El-Hoseiny MD , Hanan Ali Taha MD , Nilly Helmy Mohamed MD and Sara Ahmed Sayed. Department of Clinical and Chemical Pathology Beni Suef Teaching Hospital, Faculty of Medicine, BENI-SUEF UNIVERSITY Egypt. Department of Internal Medicine , Beni Suef Teaching Hospital, Faculty of Medicine, BENI-SUEF UNIVERSITY, Egypt.

Methylation of promoter region of CDX2 gene in colorectal cancer.

The incidence of colorectal cancer is on the increase owing to changes in daily diet. In the present study, the methylation status of caudal type homeobox transcription factor 2 (CDX2) gene in lesion tissue of colorectal cancer (CRC) was investigated. Additionally, the correlation between the promoter methylation of CDX2 gene, CRC and gene expression in patients with CRC and normal population was examined. Between April 2014 and May 2015 78 cases with CRC were enrolled in the study. Using methylation-specific polymerase chain reaction (PCR), the promoter methylation of CDX2 in normal tissues and colorectal tissues was examinned. Through the fluorescence quantitative PCR technique, the expression levels of CDX2 gene were determined in a normal population and lesion tissue of patients with CRC. At the same time, we evaluated the levels of the CDX2 gene product in the normal population and lesion tissue of patients with CRC. The results showed that the methylation rate of the promoter region of CDX2 gene in normal colorectal tissue was 43.5%, whereas that in the lesion tissue of CRC was 78.5%. The result was statistically significant (P<0.05). The quantity of mRNA and protein expression of CDX2 gene in colorectal and normal tissue was significantly different (P<0.05). In conclusion, the methylation of the CDX2 gene promoter region was associated with risk of CRC, i.e., methylation of the promoter region of CDX2 gene favors the occurrence of CRC.

Immunohistochemistry for CDX2 Expression in Non-Goblet-Cell Barrett’s Oesophagus

Interest has increased in CDX2 gene expression in oesophageal non-goblet-cell columnar metaplasia as recent investigations indicate such metaplasia possesses neoplastic potential. This study aims to assess expression of the transcription factor CDX2 specifically in non-goblet-cell cardia and fundic oesophageal metaplastic tissue, and to compare the location of CDX2 expression in non-goblet-cell specimens to that in goblet-cell specimens. A total of 43 patient specimens (20 fundic-type metaplasia, 42 cardia-type metaplasia and 18 intestinal metaplasia goblet cellpositive) were examined in this study. These were selected over six months from a patient database using the systematised nomenclature of human and veterinary medicine coding system (SNOMED). CDX2 was detected in patient specimens with an anti-CDX2 mouse monoclonal antibody. The types of mucosa in each specimen were confirmed by haematoxylin and eosin (H&E) staining. Fundic specimens were consistently CDX2-negative (0%). CDX2 expression was distinct in 55% of cardia and 100% of intestinal cases. Nearly all cardia-positive cases displayed focal expression (95.5%) and all intestinal cases displayed diffuse distribution of expression. Almost all cardia-and intestinal-positive specimens demonstrated epithelial expression (95.5% and 100%, respectively). The percentage of cardia-positive specimens with deep tissue expression was lower than in intestinal specimens (31.8% vs. 94.4%, respectively). This study confirms CDX2 as an early marker for Barrett’s oesophagus in the absence of goblet cells as expression was noted in cardia metaplasia. CDX2 appears to induce the transformation of the normal oesophageal mucosa to cardia type, which then differentiates to an intestinal type under the influence of gastro-oesophageal reflux disease.

Dysregulation of CDX1, CDX2 and SOX2 in patients with gastric cancer also affects the non-malignant mucosa

The interplay between gastric and intestinal transcription factors has an important impact on gastric carcinogenesis. We compared the gene expression of CDX1, CDX2, SOX2 and related downstream genes in tumour...