ABSTRACT
Abstract: Introduction Most cases of colorectal cancer (CRC) occur sporadically; however, ~3% to 6% of all CRCs are related to inherited syndromes, such as Lynch syndrome and familial adenomatous polyposis (FAP). The adenomatous polyposis coli (APC) andmutY DNA glycosylase (MUTYH) germline mutations are the main genetic causes related to colorectal polyposis. Nevertheless, in many cases mutations in these genes have not been identified. The aim of the present case report is to describe a rare case of genetic colorectal polyposis associated with the axis inhibition protein 2 (AXIN2) gene. Case Report: The first colonoscopy screening of a 61-year-old male patient with no known family history of CRC revealed ~ 50 colorectal polyps. A histological evaluation of the resected polyps showed low-grade tubular adenomas. Germline genetic testing through a multigene panel for cancer predisposition syndromes revealed a pathogenic variant in the AXIN2 gene. In addition to colorectal polyposis, the patient had mild features of ectodermal dysplasia: hypodontia, scant body hair, and onychodystrophy. Discussion: The AXIN2 gene acts as a negative regulator of the Wnt/β -catenin signaling pathway, which participates in development processes and cellular homeostasis. Further studies are needed to support the surveillance recommendations for carriers of the AXIN2 pathogenic variant. (AU)
Subject(s)
Humans , Male , Middle Aged , Adenomatous Polyposis Coli/diagnosis , Axin Protein/genetics , MutationABSTRACT
ABSTRACT Background : It is believed that the Wnt pathway is one of the most important signaling involved in gastric carcinogenesis. Aim : To analyze the protein expression of canonical and non-canonical Wnt pathways in gastric carcinoma. Method : The immunohistochemistry was performed in 72 specimens of gastric carcinomas for evaluating the expression of Wnt-5a, FZD5, GSK3β, axin, CK1, ubiquitin, cyclin D1 and c-myc. Results : There were significant differences for cytoplasm and nucleus ubiquitin for moderately and well differentiated tumors (p=0.03) and for those of the intestinal type of the Lauren classification (p=0.03). The absence of c-myc was related to Lauren's intestinal tumors (p=0.03). Expression of CK1 in the cytoplasm was related to compromised margin (p=0.03). Expression of cyclin D1 protein was more intense in male patients (p=0.03) There was no relation of the positive or negative expression of the Wnt-5a, FZD5, GSK3 and Axin with any clinicopathological variables. Conclusion: The canonical WNT pathway is involved in gastric carcinoma.
RESUMO Racional : Acredita-se que a via Wnt é uma das mais importantes da sinalização envolvidas na carcinogênese gástrica. Objetivos : Analisar a expressão das proteínas das vias Wnt canônicas e não-canônicas no carcinoma gástrico e relacionar sua expressão com as variáveisclinicopatológicas. Método : Foram coletadas 72 amostras de carcinoma gástrico, e áreas representativas do tumor foram selecionadas para o Tissue Microarray. Imunoistoquímica foi realizada para avaliar a expressão de Wnt-5a, FZD5, GSK3β, axina, CK1, ubiquitina, ciclina D1 e c-myc. Resultados : Houve diferenças significativas para a expressão de ubiquitina no citoplasma e núcleo para tumores moderadamente e bem diferenciados (p=0,03) e para aqueles do tipo intestinal da classificação de Lauren (p=0,03). A expressão negativa da proteína c-myc no citoplasma foi relacionada aos tumores intestinais de Lauren (p=0,028). A expressão positiva de CK1 no citoplasma das células neoplásicas foi relacionada a tumores com margens cirúrgicas livre de envolvimento neoplásico (p=0,03). A expressão positiva da proteína ciclina D1 foi maior nos tumores dos homens (p=0,03). Não houve relação da expressão positiva ou negativa das proteínas Wnt-5a e FZD5 no citoplasma ou núcleo com quaisquer variáveis clinicopatológicas. O mesmo foi observado para GSK3β e Axin. Conclusões : A relação da expressão das proteínas da via canônica com as variáveis epidemiológicas e tumorais sugere sua participação na carcinogênese gástrica. Por outro lado, a ausência da relação das expressões das proteínas da via não-canônica sugere sua não participação na carcinogênese gástrica.
Subject(s)
Humans , Male , Female , Stomach Neoplasms/chemistry , Carcinoma/chemistry , Wnt Signaling Pathway , Neoplasm Proteins/analysis , Reference Values , Stomach Neoplasms/pathology , Immunohistochemistry , Carcinoma/pathology , Proto-Oncogene Proteins c-myc/analysis , Cyclin D1/analysis , Ubiquitin/analysis , Casein Kinase I/analysis , Frizzled Receptors/analysis , Axin Protein/analysis , Carcinogenesis , Glycogen Synthase Kinase 3 beta/analysis , Wnt-5a Protein/analysis , Neoplasm StagingABSTRACT
<p><b>OBJECTIVE</b>To investigate the mRNA and protein expression of axin inhibitor (AXIN), β-chain protein (β-catenin), matrix metalloproteinase-7 (MMP-7) and matrix metalloproteinase-9 (MMP-9), and their relationship in lymphoma cells.</p><p><b>METHODS</b>The expressions of MMP-7, MMP-9, β-catenin and AXIN in lymphoma cell lines were detected by Western blot and RT-PCR. Moreover, the lymphoma cells with relatively low expression of AXIN were grouped and were transiently transfected by using pcDNA5-His-β-catenin and pCMV5-HA-AXIN; the protein and mRNA expression of MMP-7, MMP-9 and β-catenin in lymphoma cells was detected by Western blot and RT-PCR, respectively; the cell infiltration and migration ability in group with stable ligh expression of AXIN, group of interfering stable high expression of AXIN and blank control group were analyzed by transwell experiment.</p><p><b>RESULTS</b>The AXIN negatively correlated with MMP-7, MMP-9 and β-catenin expression in lymphoma cell lines. After the up-regulation of AXIN, the mRNA expression of MMP7, MMP-9 and β-catenin in Raji cells all not significantly changed, while the pratein expression of MMP-7, MMP-9 and β-catenin all significantly decreased (P<0.05); after the up-regulation of β-catenin, the mRNA and protein expression of MMP-7, MMP-9 was also up-regulated significantly (P<0.05). After interfering the AXIN, the mRNA expression of MMP-7, MMP-9 and β-catenin in group with stable high expression of AXIN all not changed significantly, while protein expression of MMP-7, MMP-9 and β-catenin was down-regulated significantly (P<0.05); after interfering the β-catenin, the protein and mRNA expression of MMP-7 and MMP-9 in group with stable high expression of AXIN all were down-regulated significantly(P<0.05).</p><p><b>CONCLUSION</b>The up-regulation of AXIN expression in lymphoma cells can lead to decrease of β-catenin expression and the resuts in significant decrease of MMP-7 and MMP-9 expression, there by plays a role to block the infiltration and migration of lymphoma cells.</p>
Subject(s)
Humans , Axin Protein , Lymphoma , Matrix Metalloproteinase 7 , Matrix Metalloproteinase 9 , RNA, Messenger , beta CateninABSTRACT
Abstract Non-syndromic cleft lip with or without palate (NSCL/P) is a common congenital malformation worldwide, with complex etiology. It has been proposed that interaction of genes and environmental factors play a role in the predisposition to this disease. Objectives: The aim of this study was to examine the association between AXIN2 (axis inhibition protein 2) rs7224837, BMP4 (bone morphogenetic protein 4) rs17563, and IRF6 (interferon regulatory factor 6) rs861019 and 2235371 polymorphisms and NSCL/P in an Iranian population. Material and Methods: This case-control study was carried out on 132 unrelated NSCL/P patients and 156 healthy subjects. The variants were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The findings suggest that BMP4 rs17563 polymorphism significantly decreased the risk of NSCL/P in codominant (OR=0.36, 95%CI=0.17-0.79, p=0.012, CT vs CC and OR=0.11, 95%CI=0.01-0.88, p = 0.019, TT vs CC), dominant (OR=0.30, 95%CI=0.15-0.62, p = 0.0007, CT+TT vs CC), recessive (OR=0.12, 95%CI=0.02-0.99, p = 0.023, TT vs CC+CT), overdominant (OR=0.39, 95%CI = 0.18-0.84, p=0.021, CT vs CC+TT), and allele (OR=0.28, 95%CI=0.15-0.55, p<0.0001, T vs C) inheritance models. Our findings did not support an association between AXIN2 rs7224837 and IRF6 rs861019 polymorphism and risk/protection of NSCL/P. The IRF6 2235371 variant was not polymorphic in our population. Conclusion: The results indicate that the BMP4 rs17563 variant is likely to confer a protective effect against the occurrence of NSCL/P in a sample of the southeast Iranian population.
Subject(s)
Humans , Male , Female , Child , Cleft Lip/genetics , Cleft Palate/genetics , Interferon Regulatory Factors/genetics , Bone Morphogenetic Protein 4/genetics , Axin Protein/genetics , Polymorphism, Restriction Fragment Length , Case-Control Studies , Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Gene Frequency , Genotype , IranABSTRACT
Familial adenomatous polyposis (FAP) is one of the most common hereditary colorectal cancers. Its intestinal and extra-intestinal manifestations are correlated with mutation sties of the APC gene. Potential gene modulation sites in patients who have typical clinical manifestations but with unidentified APC mutations are also discussed, which included MUTYH gene, AXIN gene and certain epigenetic changes. With the generalization of Precision Medicine, to offer individualized treatment and surveillance strategy based on the genotype-phenotype correlation will be of great value for FAP patients. This review focuses on the research advance in genotype - phenotype correlation studies of FAP patients.
Subject(s)
Humans , Adenomatous Polyposis Coli , Genetics , Axin Protein , Genetics , DNA Glycosylases , Genetics , Genes, APC , Genetic Association Studies , Genetic Predisposition to Disease , Mutation , beta Catenin , GeneticsABSTRACT
ABSTRACT Objective To evaluate the destruction complex of beta-catenin by the expression of the proteins beta-catetenin, adenomatous polyposis coli, GSK3β, axin and ubiquitin in colorectal carcinoma and colonic adenoma. Methods Tissue samples from 64 patients with colorectal carcinoma and 53 patients with colonic adenoma were analyzed. Tissue microarray blocks and slides were prepared and subjected to immunohistochemistry with polyclonal antibodies in carcinoma, adjacent non-neoplastic mucosa, and adenoma tissues. The immunoreactivity was evaluated by the percentage of positive stained cells and by the intensity assessed through of the stained grade of proteins in the cytoplasm and nucleus of cells. In the statistical analysis, the Spearman correlation coefficient, Student’s t, χ2, Mann-Whitney, and McNemar tests, and univariate logistic regression analysis were used. Results In colorectal carcinoma, the expressions of beta-catenin and adenomatous polyposis coli proteins were significantly higher than in colonic adenomas (p<0.001 and p<0.0001, respectively). The immunoreactivity of GSK3β, axin 1 and ubiquitin proteins was significantly higher (p=0.03, p=0.039 and p=0.03, respectively) in colorectal carcinoma than in the colonic adenoma and adjacent non-neoplastic mucosa. The immunohistochemistry staining of these proteins did not show significant differences with the clinical and pathological characteristics of colorectal cancer and colonic adenoma. Conclusions These results suggest that, in adenomas, the lower expression of the beta-catenin, axin 1 and GSK3β proteins indicated that the destruction complex of beta-catenin was maintained, while in colorectal carcinoma, the increased expression of beta-catenin, GSK3β, axin 1, and ubiquitin proteins indicated that the destruction complex of beta-catenin was disrupted.
RESUMO Objetivo Avaliar o complexo de destruição da betacatenina no carcinoma colorretal e no adenoma do colo pela expressão das proteínas betacatenina, adenomatous polyposis coli, GSK3β, axina e ubiquitina. Métodos Amostras de tecidos de 64 doentes com carcinoma colorretal e de 53 pacientes com adenoma do colo foram analisadas. Blocos de tecidos foram submetidos ao estudo imuno-histoquímico com anticorpos policlonais nos tecidos do carcinoma, mucosa não neoplásica adjacente e adenoma. A imunorreatividade foi avaliada pela porcentagem de positividade de células coradas e pela intensidade do grau de coloração das proteínas no citoplasma e no núcleo das células. Na análise estatística, foram utilizados o coeficiente de correlação de Spearman, os testes t de Student, χ2, Mann-Whitney e de McNemar, e a análise de regressão logística univariada. Resultados No carcinoma colorretal, as expressões da betacatenina e da adenomatous polyposis coli foram significativamente maiores do que em adenomas do colo (p<0,001 e p<0,0001, respectivamente). A imunorreatividade das proteínas GSK3β, axina 1 e ubiquitina foi significativamente maior (p=0,03, p=0,039 e p=0,03, respectivamente) no carcinoma colorretal do que no adenoma e na mucosa não neoplásica adjacente. A coloração imuno-histoquímica dessas proteínas não apresentou diferenças significantes em relação às características clinicopatológicas do câncer colorretal e do adenoma. Conclusões Em adenomas, as menores expressões de betacatenina, axina 1 e GSK3β indicaram que o complexo de destruição da betacatenina estava conservado, enquanto que, no carcinoma colorretal, o aumento das expressões da betacatenina, GSK3β, 1 axina, e ubiquitina indicaram que o complexo de destruição de betacatenina estava alterado.
Subject(s)
Humans , Male , Female , Middle Aged , Aged , Aged, 80 and over , Rectal Neoplasms/metabolism , Carcinoma/metabolism , Adenoma/metabolism , Colonic Neoplasms/metabolism , Axin Signaling Complex/metabolism , Neoplasm Proteins/metabolism , Rectal Neoplasms/pathology , Immunohistochemistry , Carcinoma/pathology , Adenoma/pathology , Retrospective Studies , Longitudinal Studies , Colonic Neoplasms/pathology , Adenomatous Polyposis Coli/metabolism , Ubiquitin/metabolism , beta Catenin/metabolism , Axin Protein/metabolism , Wnt Signaling Pathway , Glycogen Synthase Kinase 3 beta/metabolismABSTRACT
Delphinidin possesses strong anti-oxidant, anti-inflammatory, and anti-cancer properties. Suppression of the Wnt/β-catenin signaling pathway is a potential strategy for chemoprevention and therapy. As aberrant activation of the β-catenin signaling pathway contributes to prostate cancer progression, we evaluated the effect of delphinidin on this pathway in human PC3 prostate cancer cells. An MTT assay showed that treatment with delphinidin (15-180 μM, 72 hours) resulted in a dose-dependent growth inhibition of cells. Treatment with delphinidin increased the phosphorylation of serine or threonine residues on β-catenin and decreased the levels of cytoplasmic β-catenin. Moreover, treatment with delphinidin inhibited the nuclear translocation of β-catenin and the expression of β-catenin target genes such as cyclin D1, c-myc, Axin-2, and T cell factor-1. Delphinidin also induced the phosphorylation of glycogen synthase kinase 3β and the expression of adenomatous polyposis coli and Axin proteins. Our results indicate that inhibition of cell growth by delphinidin is mediated, at least in part, through modulation of the β-catenin signaling pathway. We suggest that delphinidin is a potent inhibitor of Wnt/β-catenin signaling in prostate cancer cells.
Subject(s)
Humans , Adenomatous Polyposis Coli , Anthocyanins , Axin Protein , beta Catenin , Chemoprevention , Cyclin D1 , Cytoplasm , Glycogen Synthase Kinases , Phosphorylation , Prostate , Prostatic Neoplasms , Serine , ThreonineABSTRACT
<p><b>OBJECTIVE</b>To investigate that the role of Axin in regulating the invasion and migration ability of lymphoma cells and explore the molecular mechanisms.</p><p><b>METHODS</b>The expressions of Axin, β-catenin, MMP7, and MMP9 were detected in different lymphoma cell lines by RT-PCR and Western blotting. A lymphoma cell line with low Axin expressions was transiently transfected with pCMV5-HA-Axin and pcDNA5-His-β-catenin plasmid, and the expressions of β-catenin, MMP7, and MMP9 mRNA and protein were observed. A lymphoma cell model stably overexpressing Axin was transfected with AXIN-shRNA and β-catenin-shRNA, and the changes in β-catenin, MMP7, and MMP9 cexpressions were observed. The changes in the invasion and migration abilities of this cell model were assessed following Axin knockdown.</p><p><b>RESULTS</b>In the lymphoma cell lines tested, the Axin expression showed a negative correlation with β-catenin, MMP7, and MMP9 expressions. In Raji cells with a low Axin expression, overexpression of Axin resulted in decreased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, and overexpression of β-catenin obviously increased MMP7 and MMP9 mRNA and protein expressions. In the cells with stable Axin overexpression, Axin knockdown caused increased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, while β-catenin knockdown caused lowered expressions of MMP7 and MMP9 and suppressed cell invasion and migration.</p><p><b>CONCLUSION</b>In lymphoma cells, Axin overexpression can decrease the expression of β-catenin, which in turn decreases the expressions of MMP7 and MMP9 to inhibit the cell invasion and migration.</p>
Subject(s)
Humans , Axin Protein , Genetics , Metabolism , Cell Line, Tumor , Down-Regulation , Gene Knockdown Techniques , Lymphoma , Genetics , Metabolism , Pathology , Matrix Metalloproteinase 7 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Neoplasm Invasiveness , Neoplasm Metastasis , RNA, Messenger , RNA, Small Interfering , Transfection , beta Catenin , MetabolismABSTRACT
Colorectal cancer is linked to several signaling pathways such as Wnt pathway. Our objective is to detect and verify the integrity of protein members of Wnt signaling pathway in colorectal carcinoma and non-neoplastic colorectal tissue. Sixty-four patients with colorectal carcinoma provided samples of colorectal neoplasia and non-neoplastic tissues, which were prepared in tissue microarray blocks and subjected to immunohistochemical analysis. The primary antibodies used were Wnt-1, Wnt-2, Wnt-5a Frizzled-1, Frizzled-5 and axin. Immunoexpression of Wnt-2 protein was significantly lower in colorectal tumor tissue and axin protein immunoexpression was significantly higher in tumor tissue. There was no significant difference in the expression of Wnt-1, Wnt-5a, Frizzled-1 and Frizzled-5 proteins in both tissues. The higher expression of Wnt-2 protein in non-neoplastic colorectal tis- sue suggests the participation during the hyperproliferative stage of colorectal mucosa. The increased axin protein immunoexpression in colorectal tumor suggests a decrease in the formation of the beta-catenin destructor complex. (AU)
O câncer colorretal está ligado a várias vias de sinalização, como a via Wnt. Nosso objetivo é detectar e verificar a integridade das proteínas da via de sinalização Wnt no carcinoma colorretal e no tecido colorretal não neoplásico. Sessenta e quatro pacientes com carcinoma colorretal forneceram amostras de neoplasia e tecidos não neoplásicos, que foram colocadas em blocos de tissue microarray e submetidas à análise imuno-histoquímica. Os anticorpos primários utilizados foram Wnt-1, Wnt-2, Wnt-5a Frizzled-1, Frizzled-5 e axina. A imunoexpressão da proteína Wnt-2 foi significativamente menor no tecido tumoral e a imunoexpressão da proteína axina foi significativamente superior no tecido do tumor. Não houve diferença significativa na expressão de Wnt-1, Wnt-5a, frizzled-1 e nas proteínas Frizzled 1 e 5 em ambos os tecidos. A maior expressão de Wnt-2 da proteína no tecido colorretal não neoplásico sugere a participação desta proteína durante o estágio de hiperproliferação da mucosa colorretal. O aumento da imunoexpressão da proteína axina no tumor colorretal sugere uma diminuição na formação do complexo de destruição da proteína beta-catenina. (AU)
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Rectal Neoplasms , Colorectal Neoplasms , Wnt Proteins , Intestinal Mucosa/immunology , Frizzled Receptors , Axin Protein , Intestinal Mucosa/pathologyABSTRACT
OBJECTIVE: To assess the effect of Leisure-time physical activity (LTPA) on cardiometabolic risk by nutritional status in Mexican children and adolescents. METHODS: This was a cross-sectional study conducted with 1,309 participants aged between 5 and 17 years. Nutritional status was classified according to the BMI Z-score by age and gender. A previously validated questionnaire was used to evaluate LTPA; a cardiometabolic risk score was calculated. Multiple linear regression analysis was performed to assess the effect of LTPA on cardiometabolic risk. RESULTS: After adjusting for risk factors, mild LTPA were positively associated with cardiometabolic risk score (ßMildvsIntenseLTPA: 0.68; 95% CI: 0.18 to 1.18; pfortrend = 0.007). This association became stronger when estimated for overweight (ß MildvsIntenseLTPA: 1.24; 95% CI: 0.24 to 2.24; pfortrend = 0.015) and obese participants (ß MildvsIntenseLTPA: 1.02; 95% CI: 0.07 to 1.97; pfortrend= 0.045) CONCLUSION: Mild LTPA was positively associated with cardiometabolic risk in overweight and obese children and adolescents. Given the emerging childhood obesity epidemic in Mexico, these results may be useful in the design of strategies and programs to increase physical activity levels in order to achieve better health. .
OBJETIVO: Avaliar o efeito da prática de AFL sobre o risco cardiometabólico em crianças e adolescentes mexicanos de acordo com sua situação nutricional. MÉTODOS: Estudo transversal feito com 1.309 participantes de cinco a 17 anos. A situação nutricional foi classificada de acordo com o escore z de IMC por idade e sexo. Um questionário validado anteriormente foi usado para avaliar a AFL; foi calculado um escore de risco cardiometabólico. A análise de regressão linear múltipla foi feita para avaliar o efeito de AFL sobre o risco cardiometabólico. RESULTADOS: Após o ajuste de acordo com os fatores de risco, a AFL leve foi positivamente associada ao escore de risco cardiometabólico (ßAFLLevexIntensa: 0,68; IC 95%: 0,18 a 1,18; p paratendência = 0,007). Essa associação foi mais intensa quando estimada para participantes acima do peso (ßAFLLevexIntensa: 1,24; IC 95%: 0,24 a 2,24; p paratendência = 0,015) e obesos (ßAFLLevexIntensa: 1,02; IC 95%: 0,07 a 1,97; p paratendência = 0,045). CONCLUSÃO: A AFL leve foi positivamente associada ao escore de risco cardiometabólico em crianças e adolescentes acima do peso e obesos. Considerando a epidemia de obesidade infantil emergente no México, esses resultados poderão ser úteis na elaboração de estratégias e programas para aumentar os níveis de atividade física a fim de obter uma saúde melhor. .
Subject(s)
Animals , Humans , Mice , Axin Protein/genetics , Colorectal Neoplasms/genetics , DNA-Binding Proteins/genetics , Lymphoid Enhancer-Binding Factor 1/genetics , Tankyrases/antagonists & inhibitors , Transcription Factors/genetics , beta Catenin/genetics , Cell Line , Cell Line, Tumor , Signal Transduction/genetics , Transcription, Genetic/genetics , Wnt Proteins/geneticsABSTRACT
Background: In 2007, a Clinical-Case-Portfolio (CCP) was introduced as a new assessment instrument for fourth grade undergraduate medical students. Since then, several changes have been implemented such as reduction on the number of clinical cases, peer review and the introduction of virtual patient to the portfolio. Aim: To describe the virtual patient model incorporated to the CCP and assess the perception of this change and its effects on the performance of undergraduate students. Material and Methods: Virtual patients were implemented based on prototype clinical cases with specific syndromes. Students perceptions about CCP before and after the introduction of virtual patients were evaluated using a validated questionnaire that was answered voluntarily and anonymously. Results: Overall perception of CCP significantly improved after the incorporation of virtual patients (97.1 ± 24.9 and 111.3 ± 25.7 points; 57.8 and 66.2% respectively). The same improvements were observed for the domains Student Learning, Organization and Evaluation, Teaching Methodology and Integration. In both years, students obtained high grades in CCP evaluations. However CCP grades were not significantly correlated with integrated final grades. Conclusions: The incorporation of virtual patients improved undergraduate students perception of CCP.
Subject(s)
Animals , Mice , Apoptosis , Axin Protein/metabolism , Enzyme Activation , Poly(ADP-ribose) Polymerases/metabolism , Protein Serine-Threonine Kinases/antagonists & inhibitors , Adenosine Triphosphate/metabolism , Apoptosis Inducing Factor/genetics , Apoptosis Inducing Factor/metabolism , Aurora Kinases , Cell Line , Cell Membrane/metabolism , Cell Membrane/physiology , Mitochondria/metabolism , Protein Kinase Inhibitors/pharmacology , RNA Interference , Time-Lapse ImagingABSTRACT
<p><b>OBJECTIVE</b>To discuss the changes in Wnt pathway inhibiting factors in esophageal precancerosis lesions induced by methyl benzyl nitrosamine (MBNA) and the effect of Gexia Zhuyu decoction.</p><p><b>METHOD</b>Wistar rats were subcutaneously injected with MBNA (3.5 mg x kg(-1) for twice per week to establish the model. Since the 1st day after the model establishment, they were orally administered with Gexia Zhuyu decoction (16, 8 mg x kg(-1)). At the 10th week, esophageal tissues were collected to observe the pathological changes of esophageal mucosa, detect SFRP1, sFRP4, Axin1, Axin2 and GSK-3β mRNA levels.by fluorescent quantitation PCR analysis and β-catenin protein level by Western blotting.</p><p><b>RESULT</b>Being induced by MBNA, rats in the model group showed slight atypical hyperplasia in the histopathological examination. Compared with the normal group, Gexia Zhuyu decoction dose high and low groups showed no significant pathomorphological and histological changes. The model group showed lower gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and higher β-catenin protein expression level (P < 0.01) than the normal control group. The Gexia Zhuyu decoction low dose group showed higher gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and lower β-catenin protein expression level (P < 0.01) than the normal control group.</p><p><b>CONCLUSION</b>Up-regulated β-catenin protein level and down-regulated Wnt pathway could enhance Wnt pathway activity of MBNA-induced esophageal precancerous lesions. Gexia Zhuyu decoction could down-regulate the β-catenin protein level and up-regulate the transcription level of Wnt pathway inhibiting factors, but could not block MBNA-induced esophageal precancerosis lesions.</p>
Subject(s)
Animals , Humans , Male , Rats , Axin Protein , Genetics , Metabolism , Drugs, Chinese Herbal , Esophageal Diseases , Drug Therapy , Genetics , Metabolism , Pathology , Glycogen Synthase Kinase 3 , Genetics , Metabolism , Glycogen Synthase Kinase 3 beta , Necrosis , Nitrosamines , Proteins , Genetics , Metabolism , Rats, Wistar , Wnt Proteins , Genetics , Metabolism , Wnt Signaling PathwayABSTRACT
The Wnt signaling pathway plays crucial roles during embryonic development, whose aberration is implicated in a variety of human cancers. Axin, a key component of canonical Wnt pathway, plays dual roles in modulating Wnt signaling: on one hand, Axin scaffolds the "β-catenin destruction complex" to promote β-catenin degradation and therefore inhibits the Wnt signal transduction; on the other hand, Axin interacts with LRP5/6 and facilitates the recruitment of GSK3 to the plasma membrane to promote LRP5/6 phosphorylation and Wnt signaling. The differential assemblies of Axin with these two distinct complexes have to be tightly controlled for appropriate transduction of the "on" or "off" Wnt signal. So far, there are multiple mechanisms revealed in the regulation of Axin activity, such as post-transcriptional modulation, homo/hetero-polymerization and auto-inhibition. These mechanisms may work cooperatively to modulate the function of Axin, thereby playing an important role in controlling the canonical Wnt signaling. In this review, we will focus on the recent progresses regarding the regulation of Axin function in canonical Wnt signaling.
Subject(s)
Animals , Humans , Axin Protein , Chemistry , Metabolism , Epigenesis, Genetic , MicroRNAs , Genetics , Metabolism , Neoplasms , Genetics , Protein Processing, Post-Translational , Wnt Signaling Pathway , GeneticsABSTRACT
Our research aimed to look into the clinical traits and genetic mutations in sporadic non-syndromic anodontia and to gain insight into the role of mutations of PAX9, MSX1, AXIN2 and EDA in anodontia phenotypes, especially for the PAX9. Material and Methods The female proband and her family members from the ethnic Han families underwent complete oral examinations and received a retrospective review. Venous blood samples were obtained to screen variants in the PAX9, MSX1, AXIN2, and EDA genes. A case-control study was performed on 50 subjects with sporadic tooth agenesis (cases) and 100 healthy controls, which genotyped a PAX9 gene polymorphism (rs4904210). Results Intra-oral and panoramic radiographs revealed that the female proband had anodontia denoted by the complete absence of teeth in both the primary and secondary dentitions, while all her family members maintained normal dentitions. Detected in the female proband were variants of the PAX9 and AXIN2 including A240P (rs4904210) of the PAX9, c.148C>T (rs2240308), c.1365A>G (rs9915936) and c.1386C>T (rs1133683) of the AXIN2. The same variants were present in her unaffected younger brother. The PAX9 variations were in a different state in her parents. Mutations in the MSX1 and EDA genes were not identified. No significant diferences were found in the allele and genotype frequencies of the PAX9 polymorphism between the controls and the subjects with sporadic tooth agenesis. Conclusions These results suggest that the association of A240P with sporadic tooth agenesis still remains obscure, especially for different populations. The genotype/phenotype correlation in congenital anodontia should be verified. .
Subject(s)
Female , Humans , Male , Anodontia/genetics , Genetic Predisposition to Disease , PAX9 Transcription Factor/genetics , Polymorphism, Genetic/genetics , Axin Protein/genetics , Case-Control Studies , China , Ectodysplasins/genetics , Gene Frequency , Genetic Association Studies , MSX1 Transcription Factor/genetics , Pedigree , Radiography, Panoramic , Retrospective StudiesABSTRACT
OBJECTIVES: We investigated four components of the Wnt signaling pathway in medulloblastomas. Medulloblastoma is the most common type of malignant pediatric brain tumor, and the Wnt signaling pathway has been shown to be activated in this type of tumor. METHODS: Sixty-one medulloblastoma cases were analyzed for β-catenin gene (CTNNB1) mutations, β-catenin protein expression via immunostaining and Wnt signaling pathway-related gene expression. All data were correlated with histological subtypes and patient clinical information. RESULTS: CTNNB1 sequencing analysis revealed that 11 out of 61 medulloblastomas harbored missense mutations in residues 32, 33, 34 and 37, which are located in exon 3. These mutations alter the glycogen synthase kinase-3β phosphorylation sites, which participate in β-catenin degradation. No significant differences were observed between mutation status and histological medulloblastoma type, patient age and overall or progression-free survival times. Nuclear β-catenin accumulation, which was observed in 27.9% of the cases, was not associated with the histological type, CTNNB1 mutation status or tumor cell dissemination. The relative expression levels of genes that code for proteins involved in the Wnt signaling pathway (CTNNB1, APC, AXIN1 and WNT1) were also analyzed, but no significant correlations were found. In addition, large-cell variant medulloblastomas presented lower relative CTNNB1 expression as compared to the other tumor variants. CONCLUSIONS: A small subset of medulloblastomas carry CTNNB1 mutations with consequent nuclear accumulation of β-catenin. The Wnt signaling pathway plays a role in classic, desmoplastic and extensive nodularity medulloblastoma variants but not in large-cell medulloblastomas.
Subject(s)
Adult , Child , Female , Humans , Male , Adenomatous Polyposis Coli Protein/analysis , Axin Protein/analysis , Cerebellar Neoplasms/pathology , Medulloblastoma/pathology , beta Catenin/analysis , Adenomatous Polyposis Coli Protein/metabolism , Axin Protein/metabolism , Chi-Square Distribution , Cerebellar Neoplasms/genetics , Cerebellar Neoplasms/metabolism , Disease-Free Survival , Gene Expression , Medulloblastoma/genetics , Medulloblastoma/metabolism , Real-Time Polymerase Chain Reaction , Statistics, Nonparametric , Wnt Signaling Pathway , beta Catenin/metabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate AXIN1-related CSRNP1 gene expression and the mechanism of its transcriptional regulation in TGF-β1-induced tumor cells.</p><p><b>METHODS</b>Human lung carcinoma A549 cells or human prostate cancer PC3 cells were treated with TGF-β1 at different doses (0, 20, 40, and 80 ng/ml) or at 20 ng/ml for 0, 8, 12, or 24 h, and the dose and time effect of TGF-β1 on CSRNP1 mRNA expression in the tumor cells were evaluated with real-time RT-PCR. A549 cells were also treated with TGF-β1 and cycloheximide to clarify whether CSRNP1 expression induced by TGF-β1 required de novo protein synthesis. A549 cells transfected with pcDNA3.1, flag-SMAD3, or flag-SMAD3-mu, after serum starvation, were treated with or without TGF-β1 (20 ng/mL) for 24 h, and the overexpression of wild-type SMAD3 and dominant negative SMAD3-mu mutant were confirmed by Western blotting. The effect of SMAD3 or SMAD3-mu overexpression on CSRNP1 mRNA expression was also measured by real-time RT-PCR.</p><p><b>RESULTS</b>In both A549 and PC3 cells, TGF-β1 dose- and time-dependently stimulated CSRNP1 expression, which required de novo protein synthesis in A549 cells. Overexpression of wild-type SMAD3 significantly increased the expression of CSRNP1 mRNA induced by TGF-β1, while overexpression of dominant negative SMAD3 mutant remarkably reduced CSRNP1 mRNA expression in response to TGF-β1 in A549 cells.</p><p><b>CONCLUSION</b>TGF-β1 may contribute to CSRNP1 expression through SMAD3 activation and downstream signaling in tumor cells.</p>
Subject(s)
Humans , Apoptosis Regulatory Proteins , Genetics , Metabolism , Axin Protein , Genetics , Metabolism , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , RNA, Messenger , Genetics , Signal Transduction , Smad3 Protein , Genetics , Metabolism , Transfection , Transforming Growth Factor beta1 , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the association of Axis inhibitor-2 (AXIN2) gene rs2240308, rs8081536 and rs9913621 single nucleotide polymorphisms (SNPs) with Hirschsprung disease(HSCR).</p><p><b>METHODS</b>DNA was extracted from 120 HSCR patients and 120 healthy controls. The AXIN2 gene exon2-rs2240308, exon5- rs8081536 and exon6-rs9913621 were amplified by polymerase chain reaction (PCR). SNPs of AXIN2 gene were analyzed by restrictive endonuclease digestion with CviJI, DdeI and BstNI and DNA sequencing. The allele and genotype frequencies and risk factors of HSCR and control group were analyzed by Chi-square test.</p><p><b>RESULTS</b>No significant differences were found in genotype frequencies of CC and CT in AXIN2 rs8081536 between HSCR patients and the control group (P> 0.05). The frequencies of genotypes GG, AG and AA as well as alleles A and G genotypes in AXIN2 gene rs2240308 locus were found to be associated with HSCR (P< 0.05). The disease risk of genotypes GG and AA and allele G with was 2.091, 0.846 and 1.703, respectively. The frequencies of genotypes CC, CT and TT as well as alleles C and T in AXIN2 gene rs9913621 locus were also associated with HSCR (P< 0.05). The disease risk of the genotypes CC and TT and the allele T was 0.535, 1.113 and 1.569, respectively. Heterozygote mutation for rs2240308 was found in the HSCR patients, i.e. the GCA to CCA mutation at position 301. Heterozygosity for rs9913621 was observed in the HSCR patients, i.e. the CAC to CAG mutation at position 199.</p><p><b>CONCLUSION</b>The rs8081536 allelic variation in AXIN2 gene does not contribute to the susceptibility of HSCR in the patients. AXIN2 rs2240308 and rs9913621 allelic variation might be related to HSCR. Individuals having allele G and T in these loci are at relatively high risk for HSCR.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Axin Protein , Base Sequence , Case-Control Studies , Cytoskeletal Proteins , Genetics , DNA Mutational Analysis , Exons , Genetics , Gene Frequency , Genetic Predisposition to Disease , Genotype , Hirschsprung Disease , Genetics , Polymerase Chain Reaction , Polymorphism, Single NucleotideABSTRACT
<p><b>OBJECTIVE</b>To investigate the protein expression of Axin and beta-catenin, the exon 3 mutation status of beta-catenin and their clinicopathological correlations in non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>A total of 100 NSCLC samples and their corresponding normal lung tissues were obtained from the patients undergoing surgery in the First Affiliated Hospital of China Medical University between 2001 and 2003. Protein expressions of Axin and beta-catenin were detected by immunohistochemistry. DNA sequence alterations of exon 3 of beta-catenin were investigated by polymerase chain reaction (PCR) and direct sequencing.</p><p><b>RESULTS</b>A reduced membranous expression rate of beta-catenin was observed in 80.0% of the cases (80/100) along with a nuclear expression rate of 26.0% (26/100). There was a significant difference in beta-catenin expression between well and poorly differentiated NSCLCs. Well to moderately differentiated NSCLCs showed a reduced expression rate of 70.0% (35/50), in contrast to 90.0% (45/ 50) in poorly differentiated tumors (P = 0.012). Reduced beta-catenin expression rate was 87.3% (48/55) in cases with lymph node metastasis, in contrast to 71.1% (32/45) in cases without lymph node metastasis (P = 0.044). The positive expression rate of Axin was 48.0% (48/100). Well to moderately differentiated NSCLCs demonstrated a 60.0% positive expression rate of Axin (30/50), much higher than poorly differentiated tumors [36.0% (18/50), P = 0.016]. The positive expression rate of Axin in beta-catenin nuclear expressed NSCLCs was 15.4% (4/26), much lower than cases without beta-catenin nuclear expression [59.5% (44/74), P < 0.001]. Axin nuclear expression was found in two cases in this study, suggesting that it may function as a nuclear-cytoplasmic shuttling protein. PCR and direct sequencing failed to reveal any exon 3 mutation of beta-catenin gene.</p><p><b>CONCLUSIONS</b>The reduced membranous expression of beta-catenin is associated with poorly differentiated and lymph node positive NSCLCs. The expression of Axin is inversely correlated with the degree of tumor differentiation and nuclear expression of beta-catenin. The exon 3 mutations do not contribute to the abnormal protein expression of beta-catenin in NSCLCs.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Axin Protein , Carcinoma, Non-Small-Cell Lung , Metabolism , Pathology , Cell Differentiation , Exons , Gene Expression Regulation, Neoplastic , Lung Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Mutation , Neoplasm Staging , Repressor Proteins , Metabolism , beta Catenin , Genetics , MetabolismABSTRACT
The purpose of the present study was to observe the expression of Axin protein during cardiac remodeling in rats. Cardiac remodeling animal models were prepared with the methods of jugular venous norepinephrine (NE)-infusion or arterial-vein fistula (AVF). The ultrasonic parameters of rat hearts were recorded before sacrifice. The expressions of Axin protein were determined by Western blot in rat hearts from different remodeling models as well as cultured cardiac fibroblasts from adult rats. Cardiac concentric hypertrophy and fibrosis was induced by 3-day jugular vein infusion of NE in rats. The expression of Axin in the left ventricles increased significantly compared with that of the control group. Cardiac eccentric hypertrophy without fibrosis was induced by A-V fistula for one week in rats, and no change in Axin protein expression in the left ventricles was observed. In cultured adult rat cardiac fibroblasts, NE treatment for 24 h increased significantly the Axin protein level. It is therefore concluded that Axin protein was expressed in rat heart and increased significantly in left ventricles during NE-induced rat cardiac remodeling, which may be relevant to cardiac fibrosis.