Aspectos histológicos, genéticos y moleculares de las lesiones preneoplásicas de la mucosa gástrica / Histological, genetic and molecular aspects of preneoplastic lesions of the gastric mucosa

Siendo el cáncer gástrico la 3ª causa de muerte por cáncer en Chile, y existiendo estrategias de tamizaje consistentes en pesquisa de lesiones preneoplásicas de la mucosa gástrica, es relevante conocer los aspectos genéticos y moleculares que puedan ser aplicados, en la optimización de dichas estrategias a grupos de mayor riesgo. El objetivo de este manuscrito fue revisar la evidencia actual en los aspectos señalados, y de la inmunohistoquímica de 4 marcadores (p53, CDX2, MUC2 y S100A9) en la mucosa gástrica normal y en las lesiones preneoplásicas de la misma.

SUMMARY: Since gastric cancer is the 3rd leading cause of death from cancer in Chile, and there are screening strategies consisting of screening for preneoplastic lesions of the gastric mucosa, it is important to know certain genetic and molecular aspects that can be applied in optimizing these strategies for higher risk groups. The aim of this manuscript was to review the current evidence on the aforementioned aspects, and on the immunohistochemistry of 4 markers (p53, CDX2, MUC2 and S100A9) in normal gastric mucosa and in its preneoplastic lesions.

Differential expression of CK7, CK20, CDX2 in intestinal and pancreatobiliary types of preriampullary carcinoma

Context: Incidence of periampullary carcinoma is low, approximately 0.5–2% of all gastrointestinal malignancies. Histologic subtyping has a prognostic bearing. The purpose of this study is to differentiate periampullary carcinomas based on immunohistochemistry (IHC) by using cytokeratin 7 (CK7), cytokeratin 20 (CK20), caudal type homeobox 2 (CDX2). Aims: To analyze the usefulness of IHC as single/panel of markers that included CK7, CK20, and CDX2. Settings and Design: This was a prospective study done from January 2017 to September 2018. Subjects and Methods: A total 50 pancreaticoduodenectomy specimens were evaluated and classified as intestinal (INT) and pancreaticobiliary (PB) types based on their morphological and immunohistochemical features, respectively. The morphologic subtypes, expression of IHC markers were correlated with different histologic parameters. Statistical Analysis: Chi-square test was used to study the association between different IHC markers with histologic parameters. Probability (P) values <0.05 were regarded as statistically significant. Results: The expression of CK7, CK20, CDX2 were studied in 50 cases to classify them as INT and pancreatobiliary subtypes. CK7 has high sensitivity (88.2%), CDX2 has high specificity (96.4%), CK20+/CDX2+ has both high sensitivity (94.2 percent) and specificity (89.2 percent) in differentiating INT from pancreatobiliary subtypes. The morphologic subtypes showed correlation with two variables (tumor grade, pathologic T stage). CK20 and CK20/CDX2 expression showed a positive correlation with tumor grade, pathologic T staging, and lymphovascular invasion. Conclusions: In conclusion, morphological classification can significantly discriminate histologic types, IHC plays a moderate role. However, the combined expression of CK20 and CDX2 is helpful in subtyping.

Wnt3a but not CDX-2 expression is associated with differentiated thyroid cancer

SUMMARY OBJECTIVE: Thyroid neoplasm incidence has increased worldwide, mostly due to the advancements in medical imaging and screening rates. The aberrant Wnt/β-catenin pathway has been identified as a key mechanism, and it has also been related to the metastatic activity of differentiated thyroid cancer. We aimed to verify the difference in the expression of Wnt3a, a canonical activator of the β-catenin signaling, and CDX-2, a transcription factor upregulated by Wnt/β-catenin pathway, in multinodular goiter and differentiated thyroid cancer and to determine their prognostic value. METHODS: We included 194 thyroid tissue surgical specimen and their clinicopathological data: study group (differentiated thyroid cancer, n=154) and control group (multinodular goiter, n=40). Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded tissue by the primary antibodies Wnt3a and CDX-2. RESULTS: High Wnt3a expression was significantly associated with differentiated thyroid cancer (p=0.031). CDX-2 was negative in all differentiated thyroid cancer cases (100%) and also in multinodular goiter. Wnt3a expression was significantly associated with tumors ≤20 mm (p=0.044) and with the absence of capsule invasion (p=0.031). The multivariate analyses suggested that older age (≥55), independent of capsular invasion and tumor size, was an independent prognostic factor for Wnt3a expression (p=0.058). CONCLUSIONS: Wnt3a expression but not CDX-2 is correlated with differentiated thyroid cancer samples in comparison to multinodular goiter. Although its prognostic value was limited to tumor size and capsule invasion, a combined model in a panel of immune markers can add accuracy in the classification of challenging thyroid follicular-derived lesions.

Marcação imunoistoquímica de CDX2 e Ki67 no adenocarcinoma gástrico / CDX2 and Ki67 immunohistochemistry marking in gastric adenocarcinoma

Introdução: A proteína CDX2 é um fator de transcrição específico do intestino, que está presente no tecido gástrico apenas quando há uma metaplasia intestinal. A metaplasia intestinal é uma lesão precursora do adenocarcinoma gástrico. O Ki67 é um biomarcador de proliferação celular. Objetivo: Verificar a presença da proteína CDX2 no adenocarcinoma gástrico. Comparar a expressão da CDX2 entre os diferentes graus de diferenciação e entre os níveis de proliferação celular. Método: A partir de 62 blocos histológicos contendo amostras de adenocarcinoma gástricos (4 bem diferenciados, 30 moderadamente diferenciados e 28 pouco diferenciados), foi feita a construção de blocos multiamostrais (TMA). Procedeu-se a marcação imunoistoquímica com os anticorpos escolhidos e realizou-se a leitura da área positiva imunocoradas. Resultados: 31 amostras foram positivas para a CDX2 e 31 negativas, sem diferença significativa entres os graus de diferenciação (p = 0,576). 38 amostras foram classificadas como de baixo grau de proliferação celular e 24 como de alto grau. Não houve diferença estatística de grau de proliferação celular entre os graus de diferenciação (p = 0,676). O grau de proliferação celular variou dependendo da expressão da CDX2 (p = 0,036). Conclusão: A expressão da proteína CDX2 esteve presente em 50% dos adenocarcinomas gástricos. Não houve diferença estatística da expressão do CDX2 entres os graus de diferenciação do adenocarcinoma gástrico. A proliferação celular variou dependendo da expressão da CDX2, havendo um maior nível de proliferação celular nas amostras que apresentaram expressão positiva para CDX2

Background: CDX2 protein is an intestinal specific transcription factor that is present in the gastric tissue only when there is intestinal metaplasia. Intestinal metaplasia is a gastric adenocarcinoma precursor injury. Ki67 is a cell proliferation biomarker. Objective: Verify the presence of CDX2 protein in gastric adenocarcinoma. Compare the CDX2 expression between the differentiation degree groups and between the cell proliferation degrees. Methods: It was collected 62 paraffin blocks containing the gastric adenocarcinoma samples (4 well differentiated adenocarcinoma, 30 moderately differentiated and 28 poorly differentiated). It was made the tissue microarrays blocks (TMA), so it was proceeded to immunohistochemical staining with the anti-CDX2 and the anti-Ki67 antibodies and it was accomplished the read of the positive immune-stained area. Results: 31 samples were positive for CDX2 expression, and 31 were negative. There was no statistical difference of the CDX2 expression between the differentiation degree groups (p = 0.576). 38 samples were classified as low degree of cell proliferation and 24 as high degree. There was no statistical difference of the cell proliferation degree between the differentiation degree groups (p = 0.676). The degree of cell proliferation ranged depending on the CDX2 expression (p = 0.036). Conclusion: Protein CDX2 expression was observed in 50% of gastric adenocarcinoma samples. There was no statistical difference of the CDX2 expression between the differentiation degree groups. The cell proliferation ranged depending of CDX2 expression, with a higher index of cell proliferation in the positive CDX2 expression samples

CDX2 and gastrointestinal neoplasms / 国际肿瘤学杂志

Caudal type homeobox transcription factor 2 (CDX2) is an enteric-specific nuclear transcription factor, which is mainly expressed in the small intestine and colorectum and not expressed in normal gastric mucosa. CDX2 is involved in the occurrence, development, invasion and metastasis of gastrointestinal tumors, and its expression state is related to the clinicopathological characteristics and prognosis of patients with gastrointestinal tumors. It is expected to provide a new idea for the diagnosis and treatment of gastrointestinal tumors by further studying the mechanism of CDX2 involvement in tumor genesis and development and its relationship with clinicopathological features and prognosis.

Are cdx2, beta-catenin and wnt immunomarchers useful for evaluating the chance of disease progression or evolution to death in patients with colorectal cancer? / Os imunomarcadores cdx2, beta-catenina e wnt são úteis para avaliar a chance de progressão de doença ou a evolução para óbito em pacientes com câncer colorretal?

ABSTRACT Background: Colorectal cancer (CRC) is one of the most common types of cancer in the world. Over time, intestinal epithelial cells undergo mutations that may lead to proliferative advantage and the emergence of cancer. Mutations in the beta-catenin pathway are amongst those described in the development of CRC. Aim: To verify the existence of a relation between the presence of Wnt3, beta-catenin and CDX2 in colorectal cancer samples and clinical outcomes such as disease progression or death. Method: Wnt3a, beta-catenin and CDX2 immunohistochemistry was performed on CRC tissue microarray samples (n=122), and analysis regarding the relation between biomarker expression and disease progression or death was performed. Results: No significant difference was found between the presence or absence of CDX2, beta-catenin or Wnt3a expression and clinical stage, tumor grade, disease progression or death. Conclusion: CDX2, beta-catenin and Wnt3a are not useful to predict prognosis in patients with CRC.

RESUMO Racional: O câncer colorretal (CCR) é um dos tipos mais comuns no mundo. As células epiteliais intestinais podem sofrer mutações que ocasionam vantagem proliferativa e culminam com o surgimento do câncer. Mutações da via da beta-catenina foram descritas entre as que podem ocasioná-lo. Objetivo: Verificar a existência de relação entre a expressão de Wnt3, beta-catenina e CDX2 em amostras de câncer colorretal com os eventos clínicos progressão de doença e óbito. Método: Foi realizada análise imunoistoquímica de Wnt3a, beta-catenina e CDX2 em blocos multiamostrais de CRC (n=122), e avaliada a relação entre a expressão dos biomarcadores e os desfechos progressão de doença e óbito. Resultados: Não foram encontradas diferenças significativas entre a expressão ou ausência de CDX2, beta-catenina ou Wnt3a e estádio clínico, grau de diferenciação tumoral, presença de progressão de doença ou evolução ao óbito. Conclusão: Os marcadores CDX2, beta-catenina e Wnt3a não são úteis para predizer prognóstico em pacientes com CCR.

Mutações do gene do receptor de vitamina d e níveis séricos de vitamina d em crianças com asma / Vitamin d receptor gene mutations and vitamin d serum levels in asthmatic children

RESUMO Objetivo: Verificar a relação dos polimorfismos do gene do receptor de vitamina D (RVD) com sinais clínicos e níveis de vitamina D (VD) em asmáticos. Métodos: Estudo transversal com 77 crianças de 7 a 14 anos de um ambulatório especializado, divididas em 3 grupos: asmáticos, em uso de corticoide inalatório (ICS) por mais de um ano; asmáticos sem necessidade de ICS; não asmáticos e não alérgicos (de acordo com o International Study of Asthma and Allergies in Childhood - ISAAC. Foram avaliados: espirometria, testes alérgicos, presença do polimorfismo CDX2 do promotor do RVD por reação em cadeia da polimerase (PCR) e genotipagem de polimorfismos dos éxons 2 e 3 por PCR-SSCA (single-strand conformational analysis), imunoglobulina E (IgE) total e IgE específica para ácaros e gramíneas nos três grupos estudados. Níveis de 25-hidroxivitamina D foram dosados nos asmáticos. Resultados: A média de idade foi 10,8±2,2 anos, 57% masculinos, 38 asmáticos com ICS, 22 sem ICS e 17 não asmáticos. Rinite alérgica esteve presente em 90% dos asmáticos, polimorfismo CDX2 em 23% dos asmáticos e ausente nos controles (p=0,03). Menores níveis de volume expiratório forçado no primeiro segundo (VEF1%) foram observados nos asmáticos homozigotos para CDX2 (p=0,001). Variações nas sequências dos éxons 2 e 3 não foram relacionadas com a asma ou demais testes. Deficiência ou insuficiência de VD foi diagnosticada em 98% dos asmáticos. Não houve associação entre níveis de VD e polimorfismos genéticos dos éxons 2 e 3. Conclusões: Observou-se associação positiva entre polimorfismo CDX2 em homozigoze com asma e menores valores de VEF1%. O CDX2 pode modificar a interação celular do RVD com a vitamina, bem como pode estar associado com a asma e com a dificuldade de controle da doença.

ABSTRACT Objective: To verify the relationship between polymorphisms of the vitamin D receptor gene (VDR), clinical findings, and serum vitamin D (VD) levels in asthmatics. Methods: A cross sectional study of 77 children aged 7 to 14 years old, who were attended at a specialized clinic. The children were divided into 3 groups: asthmatics who had been using inhaled corticosteroids (ICS) for more than one year; asthmatics who had not been using ICS; non-asthmatics, and children without allergies (according to the International Study of Asthma and Allergies in Childhood ­- ISAAC). Spirometry, skin prick tests, the presence of a VDR promoter CDX2 polymorphism from an allele-specific polimerase chain reaction (PCR), exons 2 and 3 polymorphisms genotyping by PCR-SSCA (single-strand conformational analysis), total immunoglobulin E (IgE) and specific IgE to mites and grass were evaluated in these three groups. Levels of 25-hydroxyvitamin D were determined in asthmatics only. Results: The mean age of the children was 10.8±2.0 years old, 57% were male, 38 were asthmatic and using ICS, 22 were asthmatic and not using ICS, and 17 were non-asthmatic. Allergic rhinitis was present in 90% of asthmatics. Homozygous CDX2 was detected in 23% of the patients and absent in the control group (p=0.03). Lower forced expiratory volume in 1 second (FEV1%) values were observed in CDX2 homozygous asthmatics (p=0.001). Variations in the exon 2 and 3 sequences were not related to asthma or the other tests. VD deficiency or insufficiency was detected in 98% of asthmatics. There was no association between VD levels and genetic polymorphisms from exons 2 and 3. Conclusions: There was a positive association between homozygous CDX2 polymorphism, asthma and lower FEV1% values. CDX2 is capable of modifying cell interaction between VDR and VD, and it could be associated with the prevalence of asthma, and the difficulty in controlling the disease.

Expresión de EDNRB y CDX2 posibles biomarcadores en progresión al cáncer cervical / EDNRB and CDX2 expression as possible biomarkers in progression to cervical cancer

RESUMEN De acuerdo a la historia natural del cáncer del cuello uterino, en donde las lesiones preneoplásicas de bajo y alto grado pueden presentar fenómenos de regresión o progresión, existe gran interés en la búsqueda de biomarcadores que permita predecir la evolución de las lesiones preneoplásicas del cérvix hacia la progresión o regresión de la enfermedad. Estos biomarcadores pudieran ser de origen genético, o epigenético que alteren la expresión de los genes y que pudieran estar asociados con la carcinogénesis en diferentes tipos de tejido humano. El objetivo del estudio fue analizar la expresión del mARN de los genes SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en muestras negativas para lesiones intraepiteliales cervicales (n=9), muestras con lesiones intraepiteliales de bajo grado (n=10) y alto grado (n=11). Se realizó análisis de expresión de los genes mencionados mediante qRT-PCR y el análisis de los datos se realizó mediante la prueba no paramétrica de ANOVA. La diferencia estadística se determinó en valores p< 0,05. Para los genes EDNRB y CDX2 se observó disminución 66,7% en las muestras sin alteraciones histológicas cervicales, comparado con una disminución en la expresión del 50% en muestras con LIEBG y para el grupo de LIEAG del 36,4% para el gen EDNRB y del 27,3% para el gen CDX2 dando una diferencia estadísticamente significativa p= 0,02. Sugiriendo que EDNRB y CDX2 podrían ser útiles como posibles biomarcadores en la carcinogénesis cervical.

ABSTRACT According into account the natural history of cervical cancer, where low- and high-grade preneoplastic lesions may present regression or progression phenomena, there is great interest in the search for biomarkers to predict the behavior of preneoplastic lesions of the cervix. These biomarkers may be of genetic origin, or epigenetics that alter the expression of genes and that may be associated with carcinogenesis in different types of human tissue. The objective of the study was to analyze the expression of the mRNA of the SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 genes in samples negative for cervical intraepithelial lesions (n = 9), low grade intraepithelial lesions (n=10) and high grade (n = 11). Expression analysis of the mentioned genes was performed using qRT-PCR and data analysis was performed using the non-parametric ANOVA test. The statistical difference was determined in values p <0.05. For the EDNRB and CDX2 genes, a 66.7% decrease was observed in the samples without cervical histological alterations, compared to a decrease in expression of 50% in LIEBG samples and 36.4% in the LIEAG group for the EDNRB gene And 27.3% for the CDX2 gene giving a statistically significant difference p = 0.02. Suggesting that EDNRB and CDX2 could be useful as potential biomarkers in cervical carcinogenesis.

The effect and underlying mechanisms of down-regulated CDX2 on migration and invasion of colon cancer cells / 西安交通大学学报(医学版)

Objective To observe the effect of down-regulated CDX2 gene on the migration and invasion abilities of colon cancer cells (SW480 and HT29) and investigate the role and mechanisms of CDX2 gene in occurrence and development of colon cancer metastasis.Methods CDX2 gene in HT29 and SW480 cells was down-regulated using lentivirus RNA interference (RNAi) vector.The interference efficiency of CDX2 was detected by qRT-PCR and Western blotting.The effect of down-regulated CDX2 expression on colon cancer cells'migration and invasion was determined by Transwell and wound heal methods.Then the effects of down-regulated CDX2 on the expressions of epithelial-mesenchymal transition (EMT)-related genes (E-cadherin,ZEB-1,Vimentin,Twist and Snail) were detected by RT-PCR and Western blotting.Results The constructed CDX2 siRNA expression vector could significantly inhibit the expression of CDX2 in HT29 and SW480 cells.Compared with those of the cells transfected with empty vector (LV-NT-shRNA) and non-transfected cells,the migration and invasion abilities of cells transfected with LV-CDX2-shRNA were markedly enhanced (P ＜ 0.05).E-cadherin expression was reduced while expressions of ZEB-1,Vimentin,Twist,and Snail were significantly increased (all P＜0.05).Conclusion Down-regulating the expression of CDX2 can induce the occurrence of EMT,thus enhancing the invasion and migration of colon cancer cells.

Expression of CDX2 and Its Relationship with Helicobacter Pylori Infection in Different Subtypes of Intestinal Metaplasia and Gastric Cancer / 现代检验医学杂志

Objective To investigate the expression of CDX2 in different subtypes of intestinal metaplasia (IM) and gastric cancer,and its relationship with Helicobacter Pylori (H.pylori) infection.Methods The expressions of CDX2 protein were detected with immunohistochemical method in 42 cases of chronic atrophic gastritis (CAG),46 cases of CAG with IM,34 cases of paracancerous IM and 50 cases ofgastric cancer.The IM was divided into three subtypes by HID-AB staining:27 cases of IM Ⅰ,23 cases of IM Ⅱ,and 30 cases of IM Ⅲ.H.pylori infection was detected with one minute rapid urease test,serum H,pylori IgG of ELISA method and HE staining in 80 caese of IM,which were divided into 46 cases of H.pylori-posi-tive groups and 34 cases of H.pylori-negative groups.Results The positive rates of H.pylori infection in IM Ⅰ,IM Ⅱ andIM Ⅲ were 66.67％,65.22％ and 43.34％,respectively,and there was no significant difference among different subtypes ofIM (x2=3.953,P＞0.05).The positive rates of CDX2 expression in IM Ⅰ,IM Ⅱ and IM Ⅲ were 85.19％,69.57％ and 36.67％,respectively,and IM Ⅲ were significant lower than IM Ⅰ,IM Ⅱ (x2 =13.899,P＜0.001;x2 =5.638,P=0.018),and there was no significant difference between IM Ⅰ and IM Ⅱ.Comparing of different types of IM group and gastric cancer group showed that the positive rates of CDX2 expression in IM Ⅰ,IM] were significant higher than in gastric cancer group (x2 =14.517,P＜0.001;x2 =5.509,P＜0.05),but there was no significant difference between IM Ⅲ and gastric cancer group (x2 =0.088,P＞0.05).The positive rates of CDX2 expression in H.pylori-positive groups was significant higher than in H.pylori-negative groups in all of.IM (76.09％ vs 44.12％,x2 8.525,P=0.004).Comparing between different subtypes of IM showed that the positive rates of CDX2 expression in H.pylori-positive groups was significant higher than in H.pylori-negative groups in IM Ⅲ (P=0.023),but there was no significant difference between IM Ⅰ and IM Ⅱ (P＞0.05).There was also no significant difference of CDX2 expression between H.pylori-positive groups and H.pylori-negative groups in gastric cancer.Conclusion H.pylori infection may affect the progression of IM and gastric carcinogenesis by affecting the expression of CDX2 in different subtypes of IM.

Effects of RNAi targeting CDX2 gene expression on biological characters of human leukemia cells and its relevant mechanisms / 天津医药

Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase-9, Bax mRNA and protein were tested by RT-PCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase-9 and Bax mRNA and protein were significantly higher than those of other two groups (P<0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the down-regulation of BCR-ABL and the up-regulation of caspase-9 and Bax.

Expression of CDX-2 and Ki-67 in different grades of colorectal adenocarcinomas.

Background: CDX2 is a caudal homeobox gene essential for intestinal differentiation and is specifi cally expressed in colorectal adenocarcinomas. Its role in colorectal carcinogenesis is not fully elucidated. Aims and Objectives: To study the expression pattern of CDX2 and Ki-67 in different grades of colorectal adenocarcinomas and to observe the relationship of their staining patterns in various tumor stages and to look for correlation if any, between Ki-67 labeling index (Ki-67 LI) and CDX2 expression. Materials and Methods: A total of 74 cases were enrolled. Detailed clinical profi le, peroperative fi ndings, histological grading and staging were noted. Immunohistochemistry for CDX2 and Ki-67 was done, and Ki-67 LI was calculated. CDX2 staining was graded semiquantitatively, and statistical analysis was done. Result: Age of presentation ranged from 20 to 75 years, and the male:female ratio was 1.83:1. There were 8, 47 and 13 cases of well, moderate and poorly differentiated adenocarcinomas, respectively. The mean Ki-67 LI of well, moderate and poorly differentiated adenocarcinomas were 14.25, 31.34 and 43.08 respectively, and their difference was statistically signifi cant, correlation was also noted with stage. CDX2 expression appeared to be stronger in poorly differentiated cases, but there was no signifi cant difference in its expression in the different grades and stages. There was no correlation between Ki-67 LI and CDX2 immunostaining pattern. The lymph node metastasis showed CDX2 positivity in all the cases. Conclusion: Expression of CDX2 does not signifi cantly change with the grade of colorectal adenocarcinomas. However, it is an important diagnostic marker in metastatic colonic lesions. The Ki-67 LI, on the other hand, showed a strong correlation with histopathological grades.

Expression and clinical significance of CDX2 gene in adult acute myeloid leukemia / 重庆医学

Objective To explore the expression and clinical significance of caudal homeobox gene CDX2 in acute myeloid leukemia(AML) patients .Methods Bone marrow (BM )and peripheral blood (PB)samples were colleted in 114 cases of donor AML patients and 56 patients undergoing chemotherapy .The CDX2 gene expression in every patient′s mononuclera cells were de‐tected by RT‐PCR .Among these patients ,19 cases were detected the gene continuous every three months .Eight healthy PB and five patients with iron deficiency anemia BM as control .Results CDX2 gene transcript levels were detectable in bone marrow mononu‐clear cells from 114 AML patients and 13 healthy donors ,but the level of gene expression was higher in AML patients(90/114 , 78 .9% ) .There was a statistically significant difference between the AML patients and normal donor (P0 .05) .Con‐clusion Higher expression level of CDX2 gene is mostly in AML patients ,but its expression has no relation with CR rate .CDX2 gene may be a prognostic molecular marker in AML patients ,and can be used to monitor the minimal residual disease of Normal chromosome karyotype AML .

Expression and its correlations of caudal type homeobox transcription factor-2, heparin-binding epidermal growth factor-like growth factor and bone morphogenetic protein-4 in Barrett's esophagus / 中华消化杂志

Objective To explore the function and significance of caudal type homeobox transcription factor-2 (CDX)-2,heparin-binding epidermal growth factor-like growth factor (HB-EGF) and bone morphogenetic protein 4 (BMP-4) of esophageal stromal tissues in the pathogenesis of Barrett's esophagus (BE).Methods A total of 116 patients were divided into groups according to gastroscopic finds and hematoxylin-eosin (H &E) staining of biopsy samples.They were divided into control group (n=29),RE group (n=32),BE group (n=35),RE treatment group (n=10) and BE treatment group (n=10).The expression of CDX 2,HB-EGF and BMP-4 in different esophageal mucosal lesions was detected by immunohistochemical staining and the changes of positive expression levels of CDX-2,HBEGF and BMP 4 were compared among groups.Variance analysis and chi-square analysis were performed to analyze the correlation among the three factors.Results The CDX-2 positive cell number of control group ((0.0±0.0)/high power field(HPF)),RE group ((43.1±10.6)/HPF),and BE group ((67.8±11.3)/HPF) increased in turn,and the differences among three groups were statistically significant (F=67.664,P＜0.01).The HBEGF positive ((6.4±1.4)/HPF,(39.4±13.5)/HPF,(55.8±13.9)/HPF) and BMP 4 positive ((0.0±0.0)/HPF,(22.6±6.4)/HPF and ((25.1± 10.3)/HPF) cell number of three groups had the same trend and the differences among three groups were statistically significant (F HB-EGF =22.925,FBMP-4 =10.463,both P＜0.01).Except the expression of BMP-4 between RE group and BE group,there were significant differences between every other two groups (LSD test,all P＜ 0.01).The expression of CDX 2,HB EGF,BMP-4 in RE treatment group ((21.7±1.7)/HPF,(16.6±5.0)/HPF and (9.2±1.0)/HPF) and BE treatment group ((51.4±8.7)/HPF,(31.0± 10.4)/HPF and (12.7±3.9)/HPF) were lower than those in RE group and BE group respectively,the differences were also statistically significant (LSD test,all P＜0.05).In RE group,the positive rate of CDX-2 (31.2％ (10/32)) was significantly lower than that of HBEGF and BMP4 (62.5％ (20/32) and 56.2％(18/32)),and the differences were statistically significant (x2=6.275 and 4.063,both P＜0.05).However in BE group,there was no statistically significant difference in the positive rate among CDX-2(85.7％(30/35)),HB-EGF (88.6％(31/35)) and BMP-4 (74.3％(26/35),all P＞0.05).Conclusions CDX-2,HB-EGF and BMP-4 may play an important role in the pathogenesis of RE and BE.HB-EGF and BMP-4 may be involved in the early episodes of BE genesis and have promotion effects on CDX-2 expression.HB-EGF and BMP-4 may be the new target in the research and treatment of BE.

Expressions of CDX2 and β-catenin in acute lymphoblastic leukemia and their correlation / 实用医学杂志

Objective To investigate the expressions of CDX2 and β-catenin in acute lymphoblastic leukemia (ALL) and their correlation. Methods Real-time PCR was used to determine the expressions of CDX2 and β-catenin mRNA in 43 de novo ALL and 30 non-malignant patients (used as control). Results The positivity rate of CDX2 mRNA expression in ALL group was 93%, but CDX2 mRNA expression couldn′t be detected in the control group (P<0.01). The mRNA expression of β-catenin could be detected in patients in both two groups, butβ-catenin mRNA expression in the ALL group was significantly higher than in the control group (P < 0.01). And mRNA expressions of CDX2 andβ-catenin were significantly correlated with WBC counts and LDH level (P<0.01). When the ALL patients acquired complete remission (CR), the mRNA expressions of CDX2 and β-catenin were significantly decreased compared with their newly diagnosed status , while disease-relapsed the mRNA expressions of CDX2 andβ-catenin were increased again. There was significantly positive correlation between CDX2 and β-catenin mRNA expressions (r = 0.835, P = 0.000). Conclusion Up-regulation of CDX2 and activation of Wnt/β-catenin pathway coexist in the ALL patients and the mRNA expressions of CDX2 and β-catenin are positively correlated.

Comparative Analysis of Gastrointestinal Microbiota Between Normal and Caudal-Related Homeobox 2 (Cdx2) Transgenic Mice

BACKGROUND/AIMS: Caudal-related homeobox 2 (Cdx2) is expressed in the human intestinal metaplastic mucosa and induces intestinal metaplastic mucosa in the Cdx2 transgenic mouse stomach. Atrophic gastritis and intestinal metaplasia commonly lead to gastric achlorhydria, which predisposes the stomach to bacterial overgrowth. In the present study, we determined the differences in gut microbiota between normal and Cdx2 transgenic mice, using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). METHODS: Twelve normal (control) and 12 Cdx2 transgenic mice were sacrificed, and the gastric, jejunal, ileac, cecal and colonic mucosa, and feces were collected. To quantitate bacterial microbiota, we used real-time qRTPCR with 16S rRNA gene-targeted, species-specific primers. RESULTS: The total numbers of bacteria in the gastric, jejunal, ileac, cecal, and colonic mucosa of the Cdx2 transgenic mice were significantly higher than those of the normal mice. The Bacteroides fragilis group and also Prevotella were not detected in the stomach of the normal mice, although they were detected in the Cdx2 transgenic mice. Moreover, the Clostridium coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, and Prevotella were not detected in the jejunum or ileum of the normal mice, although they were detected in the Cdx2 transgenic mice. The fecal microbiota of the normal mice was similar to that of the Cdx2 transgenic mice. CONCLUSIONS: Our results showed the differences in composition of gut microbiota between normal and Cdx2 transgenic mice, which may be caused by the development of gastric achlorhydria and intestinal metaplasia in Cdx2 transgenic mice.

Immunohistochemical diagnostics in stomach cancer / Монголын Анагаах Ухаан

Although stomach cancer immunohistochemistry is similar tothe immunohistochemistry of other organ, it has great impact on diagnosis and treatment, such as its ability to reveal whether the cancer is primary or metastatic and which treatment model would be more effective in individual case. Lately, CK7, CK20 and CDX-2 immunohistochemical markers are commonly used in stomach cancers. Stomach cancer prognosis is different in each patient, depending on several factors, patients’ health status, cancer cell differentiation, and cancer cell growth. To evaluate these factors,immunohistochemic al analysis is more effective and for this purpose they use Ki-67, CD 34, BCL-2, p53, Cyclin D1, andHer- 2 markers.The evaluation of HER-2 expression should be carefully carried out, as following: 1. HER-2 expression should be evaluated on minimum 5 positive stained cells. The evaluation criteria aremicroscopic magnification and cytoplasmic membrane-stained pattern. 2. Other than the membrane-stained pattern must be excluded. HER2 gene evaluation (FISH) can confirm the HER2 IHCexpression. 3. Usage of FDA approved antibody (4B5) has the advantageof increased sensitivity. 4. The algorithm for the evaluation of HER-2 expression used for breast cancer has 50% possibility of false negativity if it is used for stomach cancer. Therefore, it is needed to beevaluated with another specific algorithm. Because HER-2 2+ and 3+ cases can improve outcome with usingTrastizumab treatment.

Effect of over-expression of transcription factor CDX2 on proliferation and cell cycle of human gastric cancer cell line SGC-7901 / 中国病理生理杂志

AIM:To study the effect and the molecular mechanism of CDX 2 over-expression on the prolifera-tion, growth and cell cycle of human gastric cancer cell line SGC-7901.METHODS:The SGC-7901 cells in LV-CDX2-GFP group were transfected with the recombinant lentivirus vector LV-CDX2-GFP, the cells in LV-GFP group were trans-fected with the negative control lentiviral vector for the negative control , and the cells in blank control group were without any treatment.The cell proliferation was detected by CCK-8 assay.The cell cycle distribution was analyzed by flow cytome-try.The expression of CDX2, Bax, Bcl-2, cyclin D1 and survivin was determined by semi-quantitative RT-PCR and Wes-tern blotting .RESULTS:Compared with LV-GFP group and blank control group , the proliferation activity of the SGC-7901 cells was significantly lower (P0.05) between LV-GFP group and blank control group .CONCLUSION:Over-expression of CDX2 mediated by lentivirus inhibits the proliferation and growth of human gastric cancer SGC-7901 cells and arrestes the cell cycle at G 0/G1 phase, which may be related to down-regulation of Bcl-2, cyclin D1 and survivin and up-regulation of Bax .

Cyclooxygenase 2 in Gastric Carcinoma Is Expressed in Doublecortin- and CaM Kinase-Like-1-Positive Tuft Cells

BACKGROUND/AIMS: Doublecortin and CaM kinase-like-1 (DCAMKL1) is a marker of stem cells expressed predominantly in the crypt base in the intestine. However, DCAMKL1-positive cells have been shown to be differentiated tuft cells rather than quiescent progenitors. Tuft cells are the only epithelial cells that express cyclooxygenase 2 (COX-2) in the normal intestinal epithelium. We previously generated Cdx2-transgenic mice as model mice for intestinal metaplasia and gastric carcinoma. In the current study, we investigated the association between COX-2 and DCAMKL1 in gastric carcinoma. METHODS: We examined the association between COX-2 and DCAMKL1 expression in gastric carcinomas in clinical samples (early gastric well-differentiated adenocarcinoma) and Cdx2-transgenic mice; and the DCAMKL1-transgenic mouse stomach using immunohistochemistry and quantitative real-time polymerase chain reaction. RESULTS: The COX-2-expressing cells were scattered, not diffusely expressed, in gastric carcinomas from humans and Cdx2-transgenic mice. DCAMKL1-positive cells were also scattered in the gastric carcinomas, indicating that tuft cells could still be present in gastric carcinoma. COX-2 was expressed in DCAMKL1-positive tuft cells in Cdx2- and DCAMKL1-transgenic mouse stomachs, whereas the Sox9 transcription factor was ubiquitously expressed in gastric carcinomas, including COX-2-positive cells. CONCLUSIONS: COX-2 is expressed in DCAMKL1-expressing quiescent tuft cells in gastric carcinoma.

A simplified one-step nuclear transfer procedure alters the gene expression patterns and developmental potential of cloned porcine embryos

Various somatic cell nuclear transfer (SCNT) techniques for mammalian species have been developed to adjust species-specific procedures to oocyte-associated differences among species. Species-specific SCNT protocols may result in different expression levels of developmentally important genes that may affect embryonic development and pregnancy. In the present study, porcine oocytes were treated with demecolcine that facilitated enucleation with protruding genetic material. Enucleation and donor cell injection were performed either simultaneously with a single pipette (simplified one-step SCNT; SONT) or separately with different pipettes (conventional two-step SCNT; CTNT) as the control procedure. After blastocysts from both groups were cultured in vitro, the expression levels of developmentally important genes (OCT4, NANOG, EOMES, CDX2, GLUT-1, PolyA, and HSP70) were analyzed by real-time quantitative polymerase chain reaction. Both the developmental rate according to blastocyst stage as well as the expression levels CDX2, EOMES, and HSP70 were elevated with SONT compared to CTNT. The genes with elevated expression are known to influence trophectoderm formation and heat stress-induced arrest. These results showed that our SONT technique improved the development of SCNT porcine embryos, and increased the expression of genes that are important for placental formation and stress-induced arrest.