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1.
Clin Chim Acta ; 505: 1-5, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32070726

RESUMO

BACKGROUND: The secretor type α(1,2)fucosyltransferase gene (FUT2) is known to be rich in population-specific polymorphisms. However, genetic variations of FUT2 have not been well examined in Latin American populations in which nonsecretors are rare. METHODS: Conventional polymerase chain reactions and direct sequencing were performed to detect single nucleotide polymorphisms (SNPs) and copy number variations (CNVs) of FUT2 in Mexicans including Americans of Mexican ancestry, Puerto Ricans, Caribbeans, and Colombians. FUT2 alleles were determined by cloning into plasmids or PHASE software. The impact of uncharacterized missense SNPs on the enzyme activity were examined by transient transfection assays and estimated by several software programs. RESULTS: Three alleles, Se357, Se, and se428, were common, and the frequency of nonsecretors was relatively low in the studied populations. We also encountered several alleles specific to Africans, Europeans, and South and East Asians including a South Asian-specific sedel. In contrast to the in silico prediction, a transient expression study suggested that both of two missense SNPs, 235G > A and 304G > A, did not impair the enzyme activity. CONCLUSIONS: The allelic polymorphism of FUT2 suggests that the modern Latin American populations were formed via genetic admixture among Native Americans and populations whose ancestors migrated from other continents. In this study, we have observed a discrepancy between in silico and functional analyses for FUT2 for the first time. Therefore, experimental functional analysis is required for evaluation of SNPs of FUT2.


Assuntos
Fucosiltransferases/genética , Alelos , Grupo com Ancestrais do Continente Asiático , Simulação por Computador , Variações do Número de Cópias de DNA , Grupo com Ancestrais do Continente Europeu , Fucosiltransferases/análise , Expressão Gênica , Frequência do Gene , Humanos , Índios Sul-Americanos , América Latina/epidemiologia , Mutação de Sentido Incorreto/genética , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Nucleotídeo Único
2.
Int J Cancer ; 146(7): 1979-1992, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-31411736

RESUMO

Removal of colorectal adenomas is an effective strategy to reduce colorectal cancer (CRC) mortality rates. However, as only a minority of adenomas progress to cancer, such strategies may lead to overtreatment. The present study aimed to characterize adenomas by in-depth molecular profiling, to obtain insights into altered biology associated with the colorectal adenoma-to-carcinoma progression. We obtained low-coverage whole genome sequencing, RNA sequencing and tandem mass spectrometry data for 30 CRCs, 30 adenomas and 18 normal adjacent colon samples. These data were used for DNA copy number aberrations profiling, differential expression, gene set enrichment and gene-dosage effect analysis. Protein expression was independently validated by immunohistochemistry on tissue microarrays and in patient-derived colorectal adenoma organoids. Stroma percentage was determined by digital image analysis of tissue sections. Twenty-four out of 30 adenomas could be unambiguously classified as high risk (n = 9) or low risk (n = 15) of progressing to cancer, based on DNA copy number profiles. Biological processes more prevalent in high-risk than low-risk adenomas were related to proliferation, tumor microenvironment and Notch, Wnt, PI3K/AKT/mTOR and Hedgehog signaling, while metabolic processes and protein secretion were enriched in low-risk adenomas. DNA copy number driven gene-dosage effect in high-risk adenomas and cancers was observed for POFUT1, RPRD1B and EIF6. Increased POFUT1 expression in high-risk adenomas was validated in tissue samples and organoids. High POFUT1 expression was also associated with Notch signaling enrichment and with decreased goblet cells differentiation. In-depth molecular characterization of colorectal adenomas revealed POFUT1 and Notch signaling as potential drivers of tumor progression.


Assuntos
Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Fucosiltransferases/genética , Proteínas Oncogênicas/genética , Adenoma/genética , Adenoma/metabolismo , Adenoma/patologia , Biomarcadores Tumorais , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/patologia , Neoplasias Colorretais/metabolismo , Progressão da Doença , Fucosiltransferases/metabolismo , Humanos , Proteínas Oncogênicas/metabolismo , Reprodutibilidade dos Testes , Microambiente Tumoral
3.
Methods Mol Biol ; 2043: 25-43, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31463900

RESUMO

Metalloproteinases of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 repeats) superfamily are extensively modified with glycan moieties. Glycosylation occurs as these enzymes are trafficked through the endoplasmic reticulum (ER) and Golgi apparatus on their way to the extracellular space and includes N-linked glycosylation, O-linked fucosylation and C-linked mannosylation. This chapter focuses on O-linked fucose, which is added to properly folded thrombospondin type 1 repeats (TSRs) in the ER by protein O-fucosyltransferase 2 (POFUT2) and elongated to a Glucoseß1-3Fucose disaccharide by ß3-glucosyltransferase (B3GLCT). Knockout of POFUT2 results in embryonic lethality in mice, and inactivating mutations in B3GLCT cause Peters plus syndrome, a congenital disorder of glycosylation in humans. Addition of the disaccharide by POFUT2 and B3GLCT stabilizes folded TSRs, enhancing folding in the ER and secretion efficiency of several ADAMTS proteins from cells. Thus, POFUT2 and B3GLCT both function as an ER quality control pathway for folding of TSRs in ADAMTS proteins. In this chapter we describe in detail the methods developed to analyze secretion defects of ADAMTS proteins upon loss of either POFUT2 or B3GLCT. The methods described include creation of CRISPR/Cas9-mediated knockout cell lines of POFUT2 and B3GLCT and use of these cell lines to analyze and quantify secretion defects of ADAMTS proteins.


Assuntos
Proteínas ADAMTS/metabolismo , Fucose/metabolismo , Fucosiltransferases/genética , Galactosiltransferases/genética , Glucosiltransferases/genética , Proteínas ADAMTS/química , Animais , Retículo Endoplasmático/metabolismo , Fucosiltransferases/metabolismo , Galactosiltransferases/metabolismo , Técnicas de Inativação de Genes , Glucosiltransferases/metabolismo , Glicosilação , Complexo de Golgi/metabolismo , Células HEK293 , Humanos , Dobramento de Proteína
4.
Exp Oncol ; 41(4): 318-322, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31868335

RESUMO

This study aimed to investigate the effects of hypoxia and serum deprivation on regulation of fucosyltransferase-3 (FUT3) expression in breast cancer cells. MATERIALS AND METHODS: FUT3 expression was evaluated in T47D and MCF7 cells. Transcriptional and protein analysis was performed under hypoxia and serum deprivation conditions after 6 and 24 hours; and after 24 and 48 hours, respectively. RESULTS: In T47D cells, experimental conditions induced a significant decrease in FUT3 expression at both, transcriptional and protein levels, while in MCF7 cells the same conditions induced a significant increase of FUT3 expression. CONCLUSIONS: Regulation of FUT3 expression under hypoxic and serum deprivation conditions may be involved in the acquisition of advantages related to apoptosis resistance and metastasis promotion, according to the intrinsic differences presented by T47D and MCF7 cells.


Assuntos
Neoplasias da Mama/genética , Fucosiltransferases/genética , Regulação Neoplásica da Expressão Gênica , Apoptose , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Feminino , Humanos , Células MCF-7 , Soro/metabolismo , Hipóxia Tumoral
5.
Clin Lab ; 65(12)2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31850709

RESUMO

BACKGROUND: A case of a para-Bombay phenotype caused by a compound heterozygous mutation in the FUT1 gene was identified in this study. METHODS: We performed an agglutination examination of anti-H serum and secretor status to assess the presence of soluble blood group substances. Genotyping of ABO and FUT1 genes was also performed. RESULTS: Our results showed the presence of A and H antigens in saliva. Based on these results, the patient in the present case was diagnosed with the para-Bombay A phenotype. Direct DNA sequencing of the ABO gene indicated A1v/O1vgenotype. FUT1 gene sequence analysis revealed that the patient harbored the compound heterozygous mutation, c.881_882delTT (p.Phe294Cysfs*40) and c.658C>T (p.Arg220Cys). CONCLUSIONS: Improper identification of this phenotype may cause inappropriate transfusions because this particular blood group may be mislabeled as group O. Therefore, blood bank staff should be well trained to solve the discrepancy between cell and serum grouping in the para-Bombay phenotype.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Fucosiltransferases/genética , Mutação , Idoso , Feminino , Genótipo , Heterozigoto , Humanos , Fenótipo
6.
PLoS Genet ; 15(11): e1008497, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31747390

RESUMO

The lipopolysaccharide O-antigen structure expressed by the European Helicobacter pylori model strain G27 encompasses a trisaccharide, an intervening glucan-heptan and distal Lewis antigens that promote immune escape. However, several gaps still remain in the corresponding biosynthetic pathway. Here, systematic mutagenesis of glycosyltransferase genes in G27 combined with lipopolysaccharide structural analysis, uncovered HP0102 as the trisaccharide fucosyltransferase, HP1283 as the heptan transferase, and HP1578 as the GlcNAc transferase that initiates the synthesis of Lewis antigens onto the heptan motif. Comparative genomic analysis of G27 lipopolysaccharide biosynthetic genes in strains of different ethnic origin revealed that East-Asian strains lack the HP1283/HP1578 genes but contain an additional copy of HP1105 and JHP0562. Further correlation of different lipopolysaccharide structures with corresponding gene contents led us to propose that the second copy of HP1105 and the JHP0562 may function as the GlcNAc and Gal transferase, respectively, to initiate synthesis of the Lewis antigen onto the Glc-Trio-Core in East-Asian strains lacking the HP1283/HP1578 genes. In view of the high gastric cancer rate in East Asia, the absence of the HP1283/HP1578 genes in East-Asian H. pylori strains warrants future studies addressing the role of the lipopolysaccharide heptan in pathogenesis.


Assuntos
Infecções por Helicobacter/genética , Lipopolissacarídeos/genética , Antígenos O/genética , Neoplasias Gástricas/genética , Grupo com Ancestrais do Continente Asiático , Fucosiltransferases/genética , Fucosiltransferases/imunologia , Glucanos/genética , Glicosiltransferases/genética , Glicosiltransferases/imunologia , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/patologia , Helicobacter pylori/genética , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Humanos , /imunologia , Lipopolissacarídeos/química , Lipopolissacarídeos/imunologia , Mutagênese , Antígenos O/imunologia , Neoplasias Gástricas/epidemiologia , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/patologia
7.
J Korean Med Sci ; 34(39): e258, 2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-31602828

RESUMO

Para-Bombay phenotypes are rare blood groups that have inherent defects in producing H antigens associated with FUT1 and/or FUT2. We report the first case of para-Bombay blood type in a Southeast Asian patient admitted at a tertiary hospital in Korea. A 23-year-old Indonesian man presented to the hospital with fever and was diagnosed with a disseminated nontuberculous mycobacterium infection and anemia. During blood group typing for blood transfusion, cell typing showed no agglutination with both anti-A and anti-B reagents. Serum typing showed strong reactivity against B cells and trace agglutination pattern with A1 cells. His red blood cells failed to react with anti-H reagents. Direct sequencing of FUT1 and FUT2 revealed a missense variation, c.328G>A (p.Ala110Thr, rs56342683, FUT1*01W.02), and a synonymous variant, c.390C>T (p.Asn130=, rs281377, Se357), respectively. This highlights the need for both forward and reverse grouping.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Fucosiltransferases/genética , Grupo com Ancestrais do Continente Asiático/genética , Transfusão de Sangue , Humanos , Indonésia , Masculino , Mutação de Sentido Incorreto , Infecções por Mycobacterium não Tuberculosas/diagnóstico , República da Coreia , Análise de Sequência de DNA , Centros de Atenção Terciária , Adulto Jovem
8.
Int J Mol Sci ; 20(18)2019 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-31500188

RESUMO

Past work has shown that the protein O-fucosyltransferase 1 (POFUT1) is involved in mammal myogenic differentiation program. Pofut1 knockdown (Po -) in murine C2C12 cells leads to numerous elongated and thin myotubes, suggesting significant defects in secondary fusion. Among the few pathways involved in this process, NFATc2/IL-4 is described as the major one. To unravel the impact of POFUT1 on secondary fusion, we used wild-type (WT) C2C12 and Po - cell lines to follow Myf6, Nfatc2, Il-4 and Il-4rα expressions during a 120 h myogenic differentiation time course. Secreted IL-4 was quantified by ELISA. IL-4Rα expression and its labeling on myogenic cell types were investigated by Western blot and immunofluorescence, respectively. Phenotypic observations of cells treated with IL-4Rα blocking antibody were performed. In Po -, we found a decrease in nuclei number per myotube and a downexpression of Myf6. The observed downregulation of Nfatc2 is correlated to a diminution of secreted IL-4 and to the low level of IL-4Rα for reserve cells. Neutralization of IL-4Rα on WT C2C12 promotes myonuclear accretion defects, similarly to those identified in Po -. Thus, POFUT1 could be a new controller of myotube growth during myogenesis, especially through NFATc2/IL-4 signaling pathway.


Assuntos
Fucosiltransferases/genética , Regulação da Expressão Gênica , Interleucina-4/metabolismo , Desenvolvimento Muscular/genética , Fibras Musculares Esqueléticas/metabolismo , Fatores de Transcrição NFATC/metabolismo , Transdução de Sinais , Animais , Diferenciação Celular/genética , Linhagem Celular , Técnicas de Silenciamento de Genes , Camundongos , Fatores de Transcrição NFATC/genética , Receptores Notch/metabolismo
9.
Braz J Med Biol Res ; 52(8): e8522, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31365696

RESUMO

Pancreaticobiliary maljunction (PBM) is associated with high risk of epithelial atypical growth and malignant transformation of the bile duct or gallbladder. However, overall changes in genetic expression have not been examined in children with PBM. Genome-wide expression was analyzed using peripheral blood samples from 10 children with PBM and 15 pediatric controls. Differentially expressed genes (DEGs) were identified using microarray. Bioinformatics analysis was conducted using Gene Ontology and KEGG analyses. The top 5 in the up-regulated genes in PBM were verified with qRT-PCR. Receiver operator characteristic curve analysis was conducted to evaluate the predictive accuracy of selected genes for PBM. The microarray experiments identified a total of 876 DEGs in PBM, among which 530 were up-regulated and the remaining 346 were down-regulated. Verification of the top 5 up-regulated genes (TYMS, MYBPC1, FUT1, XAGE2, and GREB1L) by qRT-PCR confirmed the up-regulation of MYBPC1 and FUT1. Receiver operating characteristic curve analysis suggested that FUT1 and MYBPC1 up-regulation could be used to predict PBM, with the area under the curve of 0.873 (95%CI=0.735-1.000) and 0.960 (95%CI=0.891-1.000), respectively. FUT1 and MYBPC1 were up-regulated in children with PBM, and could be used as potential biomarkers for PBM.


Assuntos
Ductos Biliares/anormalidades , Proteínas de Transporte/genética , Fucosiltransferases/genética , Perfilação da Expressão Gênica , Ductos Pancreáticos/anormalidades , Regulação para Cima/genética , Neoplasias dos Ductos Biliares/etiologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Dilatação Patológica/complicações , Dilatação Patológica/congênito , Feminino , Neoplasias da Vesícula Biliar/etiologia , Humanos , Lactente , Masculino , Análise em Microsséries
11.
PLoS Pathog ; 15(7): e1007915, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31329635

RESUMO

Expression of ABO and Lewis histo-blood group antigens by the gastrointestinal epithelium is governed by an α-1,2-fucosyltransferase enzyme encoded by the Fut2 gene. Alterations in mucin glycosylation have been associated with susceptibility to various bacterial and viral infections. Salmonella enterica serovar Typhimurium is a food-borne pathogen and a major cause of gastroenteritis. In order to determine the role of Fut2-dependent glycans in Salmonella-triggered intestinal inflammation, Fut2+/+ and Fut2-/- mice were orally infected with S. Typhimurium and bacterial colonization and intestinal inflammation were analyzed. Bacterial load in the intestine of Fut2-/- mice was significantly lower compared to Fut2+/+ mice. Analysis of histopathological changes revealed significantly lower levels of intestinal inflammation in Fut2-/- mice compared to Fut2+/+ mice and measurement of lipocalin-2 level in feces corroborated histopathological findings. Salmonella express fimbriae that assist in adherence of bacteria to host cells thereby facilitating their invasion. The std fimbrial operon of S. Typhimurium encodes the π-class Std fimbriae which bind terminal α(1,2)-fucose residues. An isogenic mutant of S. Typhimurium lacking Std fimbriae colonized Fut2+/+ and Fut2-/- mice to similar levels and resulted in similar intestinal inflammation. In vitro adhesion assays revealed that bacteria possessing Std fimbriae adhered significantly more to fucosylated cell lines or primary epithelial cells in comparison to cells lacking α(1,2)-fucose. Overall, these results indicate that Salmonella-triggered intestinal inflammation and colonization are dependent on Std-fucose interaction.


Assuntos
Fímbrias Bacterianas/metabolismo , Fucose/metabolismo , Salmonella typhimurium/patogenicidade , Animais , Aderência Bacteriana , Colite/etiologia , Colite/metabolismo , Colite/microbiologia , Feminino , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/genética , Fucosiltransferases/deficiência , Fucosiltransferases/genética , Fucosiltransferases/metabolismo , Interações entre Hospedeiro e Microrganismos , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Masculino , Camundongos , Camundongos Endogâmicos CBA , Camundongos Knockout , Óperon , Salmonelose Animal/etiologia , Salmonelose Animal/metabolismo , Salmonelose Animal/microbiologia , Salmonella typhimurium/genética , Salmonella typhimurium/fisiologia
12.
EBioMedicine ; 45: 124-138, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31279780

RESUMO

BACKGROUND: Our previous study revealed that PLAGL2 or POFUT1 can promote tumorigenesis and maintain significant positive correlations in colorectal cancer (CRC). However, the mechanism leading to the co-expression and the underlying functional and biological implications remain unclear. METHODS: Clinical tumor tissues and TCGA dataset were utilized to analyze the co-expression of PLAGL2 and POFUT1. Luciferase reporter assays, specially made bidirectional promoter vectors and ectopic expression of 3'UTR were employed to study the mechanisms of co-expression. In vitro and in vivo assays were performed to further confirm the oncogenic function of both. The sphere formation assay, immunofluorescence, Western blot and qRT-PCR were performed to investigate the effect of both genes in colorectal cancer stem cells (CSCs). FINDINGS: PLAGL2 and POFUT1 maintained co-expression in CRC (r = 0.91, p < .0001). An evolutionarily conserved bidirectional promoter, rather than post-transcriptional regulation by competing endogenous RNAs, caused the co-expression of PLAGL2 and POFUT1 in CRC. The bidirectional gene pair PLAGL2/POFUT1 was subverted in CRC and acted synergistically to promote colorectal tumorigenesis by maintaining stemness of colorectal cancer stem cells through the Wnt and Notch pathways. Finally, PLAGL2 and POFUT1 share transcription factor binding sites, and introducing mutations into promoter regions with shared transcription regulatory elements led to a decrease in the PLAGL2/POFUT1 promoter activity in both directions. INTERPRETATION: Our team identified for the first time a bidirectional promoter pair oncogene, PLAGL2-POFUT1, in CRC. The two genes synergistically promote the progression of CRC and affect the characteristics of CSCs, which can offer promising intervention targets for clinicians and researchers. FUND: National Nature Science Foundation of China, the Hunan province projects of Postgraduate Independent Exploration and Innovation of Central South University.


Assuntos
Neoplasias Colorretais/genética , Proteínas de Ligação a DNA/genética , Fucosiltransferases/genética , Regiões Promotoras Genéticas/genética , Proteínas de Ligação a RNA/genética , Fatores de Transcrição/genética , Regiões 3' não Traduzidas/genética , Animais , Carcinogênese/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Camundongos , Células-Tronco Neoplásicas/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
13.
Artigo em Inglês | MEDLINE | ID: mdl-31334132

RESUMO

Thrombospondin type I repeat (TSR) domains are commonly O-fucosylated by protein O-fucosyltransferase 2 (PoFUT2), and this modification is required for optimal folding and secretion of TSR-containing proteins. The human malaria parasite Plasmodium falciparum expresses proteins containing TSR domains, such as the thrombospondin-related anonymous protein (TRAP) and circumsporozoite surface protein (CSP), which are O-fucosylated. TRAP and CSP are present on the surface of sporozoites and play essential roles in mosquito and human host invasion processes during the transmission stages. Here, we have generated PoFUT2 null-mutant P. falciparum and Plasmodium berghei (rodent) malaria parasites and, by phenotyping them throughout their complete life cycle, we show that PoFUT2 disruption does not affect the growth through the mosquito stages for both species. However, contrary to what has been described previously by others, P. berghei PoFUT2 null mutant sporozoites showed no deleterious motility phenotypes and successfully established blood stage infection in mice. This unexpected result indicates that the importance of O-fucosylation of TSR domains may differ between human and RODENT malaria parasites; complicating our understanding of glycosylation modifications in malaria biology.


Assuntos
Fucosiltransferases/metabolismo , Plasmodium berghei/enzimologia , Plasmodium berghei/crescimento & desenvolvimento , Plasmodium berghei/metabolismo , Animais , Linhagem Celular , Culicidae/parasitologia , Modelos Animais de Doenças , Fucosiltransferases/genética , Glicosilação , Humanos , Estágios do Ciclo de Vida , Malária/parasitologia , Malária/transmissão , Malária Falciparum/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Oocistos/metabolismo , Plasmodium berghei/genética , Plasmodium falciparum/genética , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Esporozoítos/enzimologia , Esporozoítos/genética , Esporozoítos/crescimento & desenvolvimento , Esporozoítos/metabolismo
14.
EBioMedicine ; 44: 563-573, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31201143

RESUMO

BACKGROUND: Endometrial stromal cell decidualization is critical for embryo implantation. Dysfunctional decidualization leads to implantation failure, miscarriage and even pregnancy associated disorders in subsequent pregnancy trimesters. Protein glycosylation is involved in many physiological and pathological processes. Protein O-fucosyltransferase 1 (poFUT1) is the key enzyme for the O-fucosylation of proteins. However, the role and mechanism of poFUT1 in human endometrial stromal cell decidualization remain elusive. METHODS: We employed immunohistochemistry to detect the level of poFUT1 in the uterine endometrium from those of the proliferative phase, secretory phase, early pregnancy women and miscarriage patients. Using human endometrial stromal cells (hESCs) and a mouse model, the underlying mechanisms of poFUT1 in decidualization was investigated. FINDINGS: The level of poFUT1 was increased in the stromal cells of the secretory phase relative to those in the proliferative phase of the menstrual cycle, and decreased in the stromal cells of miscarriage patients compared to women with healthy early pregnancies. Furthermore, we found that poFUT1 promoted hESCs decidualization. The results also demonstrated that poFUT1 increased O-fucosylation on Notch1 in hESCs, which activated Notch1 signaling pathway. Activated Notch1 (NICD), as a specific trans-factor of PRL and IGFBP1 promoters, enhanced PRL and IGFBP1 transcriptional activity, thus inducing hESCs decidualization. INTERPRETATION: Level of poFUT1 is lower in the uterine endometrium from miscarriage patients than early pregnancy women. poFUT1 is critical in endometrial decidualization by controlling the O-fucosylation on Notch1. Our findings provide a new mechanism perspective on poFUT1 in uterine decidualization that may be a useful diagnostic and therapeutic target for miscarriage. FUND: National Natural Science Foundation of China (31770857, 31670810 and 31870794). Liaoning Provincial Program for Top Discipline of Basic Medical Sciences.


Assuntos
Decídua/fisiologia , Endométrio/fisiologia , Fucosiltransferases/metabolismo , Receptor Notch1/metabolismo , Adulto , Animais , Linhagem Celular , Proliferação de Células , Feminino , Fucosiltransferases/genética , Regulação da Expressão Gênica , Glicosilação , Humanos , Imuno-Histoquímica , Ciclo Menstrual , Camundongos , Modelos Biológicos , Gravidez , Células Estromais/metabolismo
15.
Hum Mol Genet ; 28(12): 2062-2077, 2019 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-31163085

RESUMO

Glycosylation is a common post-translational modification of proteins. Glycosylation is associated with a number of human diseases. Defining genetic factors altering glycosylation may provide a basis for novel approaches to diagnostic and pharmaceutical applications. Here we report a genome-wide association study of the human blood plasma N-glycome composition in up to 3811 people measured by Ultra Performance Liquid Chromatography (UPLC) technology. Starting with the 36 original traits measured by UPLC, we computed an additional 77 derived traits leading to a total of 113 glycan traits. We studied associations between these traits and genetic polymorphisms located on human autosomes. We discovered and replicated 12 loci. This allowed us to demonstrate an overlap in genetic control between total plasma protein and IgG glycosylation. The majority of revealed loci contained genes that encode enzymes directly involved in glycosylation (FUT3/FUT6, FUT8, B3GAT1, ST6GAL1, B4GALT1, ST3GAL4, MGAT3 and MGAT5) and a known regulator of plasma protein fucosylation (HNF1A). However, we also found loci that could possibly reflect other more complex aspects of glycosylation process. Functional genomic annotation suggested the role of several genes including DERL3, CHCHD10, TMEM121, IGH and IKZF1. The hypotheses we generated may serve as a starting point for further functional studies in this research area.


Assuntos
Fucosiltransferases/genética , Glicosiltransferases/genética , Polissacarídeos/sangue , Cromatografia Líquida de Alta Pressão , Estudos de Coortes , Fucosiltransferases/sangue , Fucosiltransferases/química , Estudo de Associação Genômica Ampla , Glucuronosiltransferase/sangue , Glucuronosiltransferase/química , Glicosilação , Fator 1-alfa Nuclear de Hepatócito/sangue , Fator 1-alfa Nuclear de Hepatócito/química , Humanos , Imunoglobulina G/metabolismo , Proteínas de Membrana/metabolismo , Polimorfismo Genético , Locos de Características Quantitativas
16.
Phytochemistry ; 165: 112050, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31252202

RESUMO

In higher plants, asparagine-linked oligosaccharides (N-glycans) in glycoproteins carry unique carbohydrate epitopes, namely, a core α1,3-fucose and/or a ß1,2-xylose, which are common determinants responsible for the cross-reactivity of plant glycoproteins due to their strong immunogenicity. While these determinants and the relevant genes have been well characterized for herbaceous plants, information concerning whether many food plants cross-react with airborne pollens is not available. In this paper, we report on the characterization of a novel core α1,3-fucosyltransferase gene identified from Mangifera indica L., one of the major plants potentially related to food allergy. Based on sequence information of plant homologues, we amplified a candidate cDNA (MiFUT11) from pericarp tissue. An in vitro assay demonstrated that the recombinant MiFUT11 protein transfers a fucose unit onto both non-fucosylated and core α1,6-fucosylated oligosaccharides. A glycoform analysis using MALDI-TOF mass spectrometry showed that the introduction of the MiFUT11 cDNA increased the production of a core α1,3- and α1,6-fucosylated pauci-mannosidic oligosaccharide in Spodoptera Sf21 cells. Our findings suggest that MiFUT11 is a functional core α1,3-fucosyltransferase gene that is involved in the assembly of cross-reactive N-glycans in mango fruit.


Assuntos
Carboidratos/biossíntese , Frutas/química , Fucosiltransferases/metabolismo , Mangifera/enzimologia , Sequência de Aminoácidos , Carboidratos/genética , Carboidratos/imunologia , Frutas/imunologia , Frutas/metabolismo , Fucosiltransferases/química , Fucosiltransferases/genética , Mangifera/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência
17.
J Neurooncol ; 143(3): 405-415, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31104223

RESUMO

PURPOSE: Metastatic non-small cell lung (NSCLC) cancer represents one of the most common types of brain metastasis. The mechanisms involved in how circulating cancer cells transmigrate into brain parenchyma are not fully understood. The aim of this work was to investigate the role of fucosylated carbohydrate epitopes CD15 and sialyated CD15s in cancer adhesion to brain-derived endothelial cells and determine their influence in blood-brain barrier (BBB) disruption METHODS: Three distinct, independent methods were used to measure brain endothelial integrity and include voltohmmeter (EVOM™), impedance spectroscopy (CellZscope®) and electric cell-substrate impedance sensing system (ECIS™). Two fucosyltransferases (FUT4 and 7) responsible for CD15 and CD15s synthesis were modulated in four human cancer cell lines (three lung cancer and one glioma). RESULTS: Overexpression of CD15 or CD15s epitopes led to increase in adhesion of cancer cells to cerebral endothelial cells compared with wild-type and cells with silenced CD15 or CD15s (p < 0.01). This overexpression led to the disruption of cerebral endothelial cell monolayers (p < 0.01). Knockdown of FUT4 and FUT7 in metastatic cancer cells prevented disruption of an in vitro BBB model. Surprisingly, although the cells characterised as 'non-metastatic', they became 'metastatic' -like when cells were forced to over-express either FUT4 or FUT7. CONCLUSIONS: Results from these studies suggest that overexpression of CD15 and CD15s could potentiate the transmigration of circulating NSCLC cells into the brain. The clinical significance of these studies includes the possible use of these epitopes as biomarkers for metastasis.


Assuntos
Barreira Hematoencefálica/patologia , Neoplasias Encefálicas/secundário , Carcinoma Pulmonar de Células não Pequenas/patologia , Adesão Celular , Células Endoteliais/patologia , Fucosiltransferases/metabolismo , Neoplasias Pulmonares/secundário , Barreira Hematoencefálica/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Movimento Celular , Células Endoteliais/metabolismo , Fucosiltransferases/genética , Humanos , Antígenos CD15/genética , Antígenos CD15/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Células Tumorais Cultivadas
18.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(6): 636-638, 2019 Jun 10.
Artigo em Chinês | MEDLINE | ID: mdl-31055825

RESUMO

OBJECTIVE: To explore the molecular basis for an individual with para-Bombay phenotype of the H blood group. METHODS: Intron 5 to 3'-UTR of the ABO gene and exon 4 of the FUT1 gene were amplified with PCR and subjected to direct sequencing. Mutations of the FUT1 gene were identified by TOPO cloning sequencing. RESULTS: Direct sequencing showed that her ABO genotype was B101/O01. TOPO cloning sequencing found that this individual had three mutations of the FUT1 gene, including an heterozygous AG deletion (CAGAGAG→CAGAG) at position 547 to 552, and two C→T mutations at positions 35 (C35T) and 293 (C293T) on the other homologous chromosome. The two alleles comprised a new recombination of mutations c.35T>C and c.293C>T, and the sequence has been submitted to NCBI (No. MG597611). CONCLUSION: A novel combination of FUT1 alleles with c.35 C>T and c.293C>T has been identified in an individual with para-Bombay phenotype.


Assuntos
Sistema ABO de Grupos Sanguíneos , Fucosiltransferases/genética , Alelos , Feminino , Genótipo , Humanos , Fenótipo
19.
J Exp Clin Cancer Res ; 38(1): 200, 2019 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-31097000

RESUMO

BACKGROUND: Acute myeloid leukemia (AML) is initiated and maintained by a unique, small subset of leukemia stem cells (LSCs). LSCs are characterized by unrestricted self-renewal and contribute to the malignancy of leukemia. Aberrant protein fucosylation is associated with AML progression. However, it is still less understood that the miR-29b/Sp1/FUT4 crosstalk involved in the fucosylation-mediated LSCs malignancy in AML. METHODS: AML cell lines were sorted by magnetic microbeads to obtain the CD34 + CD38- sub-population. The key biomarkers for LSCs were identified by flow cytometry. Fucosyltransferase genes were screened by qRT-PCR, and FUT4 was focused. Effect of FUT4 on LSCs malignancy was determined by CCK8 assay, sphere formation assay, immunofluorescence staining, apoptosis and in vivo xenografts experiments. The linkage of FUT4 promoter and Sp1 was confirmed by dual-luciferase reporter gene assay. ChIP-PCR assay was used to show the directly binding of Sp1 and FUT4 promoter. Activity of Wnt//ß-catenin pathway was determined by western blot. Overall survival curves were diagrammed by Kaplan-Meier analysis. RESULTS: Here, the expressional profiles of 11 fucosyltransferase genes were different comparing LSCs and non-LSCs of KG-1a and MOLM13 cells, whereas CD34 + CD38- cells exhibited higher expression of FUT4. Functionally, alteration of FUT4 in CD34 + CD38- cells modulated LSCs malignant behaviors both in vitro and in vivo. Transcriptional inhibitor actinomycin D (Act D) or translational inhibitor cycloheximide (CHX) prevented LSCs progression, and Sp1 was identified as the efficient regulator of FUT4 transcription. Moreover, miR-29b directly affected the binding of Sp1 and FUT4 promoter region, which further mediated LSCs proliferation, apoptosis and drug-resistance through fucosylated-CD44 via activation of Wnt/ß-catenin pathway. Clinically, Sp1 and FUT4 were up-regulated and positively correlated with poor overall survival of AML patients. CONCLUSION: These data indicated that miR-29b/Sp1/FUT4 axis promoted the malignant behaviors of LSCs by regulating fucosylated CD44 via Wnt/ß-catenin pathway. Identifying LSCs surface markers and targeting LSCs were important for the development of potential therapies in AML.


Assuntos
Fucosiltransferases/genética , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , MicroRNAs/genética , Células-Tronco Neoplásicas/metabolismo , Fator de Transcrição Sp1/genética , Via de Sinalização Wnt , Animais , Biomarcadores Tumorais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Progressão da Doença , Feminino , Perfilação da Expressão Gênica , Regulação Leucêmica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Leucemia Mieloide Aguda/mortalidade , Leucemia Mieloide Aguda/patologia , Masculino , Camundongos , Interferência de RNA , Ensaios Antitumorais Modelo de Xenoenxerto
20.
J BUON ; 24(1): 61-67, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30941952

RESUMO

PURPOSE: The current study aimed to explore the effect of miR-200c on the proliferation, migration and invasion of breast cancer cells and its relevant mechanisms. METHODS: Cell counting kit-8 (CCK-8), scratch wound healing assay and Transwell assay were performed after upregulation of miR-200c to detect the capabilities of proliferation, migration and invasion of MCF-7 breast cancer cells. Also, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were carried out to determine the expression levels of fucosyltransferase-4 (FUT4) and relevant genes in PI3K/AKT signaling pathways. RESULTS: miR-200c upregulation in MCF-7 cells decreased the capabilities of proliferation, migration and invasion in MCF-7 cells. MiR-200c could regulate the level of FUT4 in MCF-7 cells, and might affect the cell proliferation, migration and invasion through PI3K/AKT signaling pathway. CONCLUSIONS: The results of this study indicated that miR-200c might serve as a new target in the diagnosis and treatment of breast cancer. MiR-200c regulated the expression of FUT4, and affected the biological behaviors of breast cancer MCF-7 cells, such as proliferation, migration and invasion.


Assuntos
Neoplasias da Mama/patologia , Movimento Celular , Proliferação de Células , Fucosiltransferases/metabolismo , MicroRNAs/genética , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Feminino , Fucosiltransferases/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Invasividade Neoplásica , Células Tumorais Cultivadas
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