Highly sensitive cell-based assay system to monitor the sialyl Lewis X biosynthesis mediated by α1-3 fucosyltransferase-VII

Miyashiro, Masahiko; Furuya, Sachiko; Fujishige, Kotomi; Sugita, Takahisa

doi:10.1016/j.bbrc.2004.09.025

Cited by 8 publications

(7 citation statements)

References 37 publications

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“…In addition to the SERPINA1 association, one of the top five associated CpG sites for all three phenotypes in both cohorts was with a CpG in the fucosyltransferase-7 (FUT7) gene (OMIM 602030), which was relatively hypomethylated in the presence of COPD and lower lung function. FUT7 is an interesting candidate for lung disease as it encodes sialyl Lewis X, a receptor component that promotes leukocyte migration to inflamed tissue (29). In the case of COPD, neutrophilic inflammation is a prominent feature, and neutrophil adhesion molecule expression has been observed to be altered in subjects with COPD (30,31).…”

Section: Discussionmentioning

confidence: 99%

Variable DNA Methylation Is Associated with Chronic Obstructive Pulmonary Disease and Lung Function

Qiu

Baccarelli

Carey

et al. 2012

Am J Respir Crit Care Med

208

196

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Rationale: Chronic obstructive pulmonary disease (COPD) is associated with local (lung) and systemic (blood) inflammation and manifestations. DNA methylation is an important regulator of gene transcription, and global and specific gene methylation marks may vary with cigarette smoke exposure. Objectives: To perform a comprehensive assessment of methylation marks in DNA from subjects well phenotyped for nonneoplastic lung disease. Methods: We conducted array-based methylation screens, using a test-replication approach, in two family-based cohorts (n ¼ 1,085 and 369 subjects). Measurements and Main Results: We observed 349 CpG sites significantly associated with the presence and severity of COPD in both cohorts. Seventy percent of the associated CpG sites were outside of CpG islands, with the majority of CpG sites relatively hypomethylated. Gene ontology analysis based on these 349 CpGs (330 genes) suggested the involvement of a number of genes responsible for immune and inflammatory system pathways, responses to stress and external stimuli, as well as wound healing and coagulation cascades. Interestingly, our observations include significant, replicable associations between SERPINA1 hypomethylation and COPD and lower average lung function phenotypes (combined P values: COPD, 1.5 3 10 223; FEV 1 /FVC, 1.5 3 10 235; FEV 1 , 2.2 3 10 240 ). Conclusions: Genetic and epigenetic pathways may both contribute to COPD. Many of the top associations between COPD and DNA methylation occur in biologically plausible pathways. This largescale analysis suggests that DNA methylation may be a biomarker of COPD and may highlight new pathways of COPD pathogenesis.Keywords: chronic obstructive pulmonary disease; epigenetics; DNA methylation; smoking Chronic obstructive pulmonary disease (COPD) is a multifactorial complex human disease of the lungs with world-wide impact (1). Genome-wide genetic association studies have identified variants associated with COPD (2, 3) and lung function (4-6), yet each identified locus explains only a small amount of the risk for COPD. Variation in the a 1 -antitrypsin (AAT) gene (SER-PINA1) is a monogenic cause of COPD, but even in the presence of AAT deficiency the development of COPD is unpredictable. COPD fits well in the common disease genetic and epigenetic hypothesis, which promulgates that epigenetic variation may be an important mediator between genetic variation and environmental exposures (7). Specifically, DNA methylation may provide further explanation for features of COPD that are not explained fully by DNA sequence variation, such as the variable susceptibility to develop lung disease in smokers, as well as the continued elevated risk for lung function decline years after smoking cessation (8). Epigenetic studies might shed new insights into the pathogenesis of COPD susceptibility and severity.Cigarette smoking is the major environmental risk factor for lung function decline and COPD. Cigarette smoking has been demonstrated to impact global hypomethylation of repetitive genomic elements (...

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Section: Discussionmentioning

confidence: 99%

Variable DNA Methylation Is Associated with Chronic Obstructive Pulmonary Disease and Lung Function

Qiu

Baccarelli

Carey

et al. 2012

Am J Respir Crit Care Med

208

196

View full text Add to dashboard Cite

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“…Similarly, we have failed to establish constitutive TIG1A and TIG1B stable clones from CC-M2, SW620, and HCT116 colon cancer cells. Therefore, the inducible GeneSwitch system, with reported low basal expression, was used in our study [18,20]. …”

Section: Discussionmentioning

confidence: 99%

“…Inducible TIG1 stable clones were established using the GeneSwitch system (Invitrogen) as previously described with minor modifications [20]. Briefly, HCT116 cells plated in 10-cm dishes were first transfected with the pSwitch expression vector (Invitrogen) that expressed the hybrid regulatory protein using lipofectamine, and then selected in hygromycin (400 μg/mL, Invitrogen)-containing medium.…”

Section: Methodsmentioning

confidence: 99%

G protein-coupled receptor kinase 5 mediates Tazarotene-induced gene 1-induced growth suppression of human colon cancer cells

Tsai

Shyu

et al. 2011

BMC Cancer

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BackgroundTazarotene-induced gene 1 (TIG1) is a retinoid-inducible type II tumour suppressor gene. The B isoform of TIG1 (TIG1B) inhibits growth and invasion of cancer cells. Expression of TIG1B is frequently downregulated in various cancer tissues; however, the expression and activities of the TIG1A isoform are yet to be reported. Therefore, this study investigated the effects of the TIG1A and TIG1B isoforms on cell growth and gene expression profiles using colon cancer cells.MethodsTIG1A and TIG1B stable clones derived from HCT116 and SW620 colon cancer cells were established using the GeneSwitch system; TIG1 isoform expression was induced by mifepristone treatment. Cell growth was assessed using the WST-1 cell proliferation and colony formation assays. RNA interference was used to examine the TIG1 mediating changes in cell growth. Gene expression profiles were determined using microarray and validated using real-time polymerase chain reaction, and Western blot analyses.ResultsBoth TIG1 isoforms were expressed at high levels in normal prostate and colon tissues and were downregulated in colon cancer cell lines. Both TIG1 isoforms significantly inhibited the growth of transiently transfected HCT116 cells and stably expressing TIG1A and TIG1B HCT116 and SW620 cells. Expression of 129 and 55 genes was altered upon induction of TIG1A and TIG1B expression, respectively, in stably expressing HCT116 cells. Of the genes analysed, 23 and 6 genes were upregulated and downregulated, respectively, in both TIG1A and TIG1B expressing cells. Upregulation of the G-protein-coupled receptor kinase 5 (GRK5) was confirmed using real-time polymerase chain reaction and Western blot analyses in both TIG1 stable cell lines. Silencing of TIG1A or GRK5 expression significantly decreased TIG1A-mediated cell growth suppression.ConclusionsExpression of both TIG1 isoforms was observed in normal prostate and colon tissues and was downregulated in colon cancer cell lines. Both TIG1 isoforms suppressed cell growth and stimulated GRK5 expression in HCT116 and SW620 cells. Knockdown of GRK5 expression alleviated TIG1A-induced growth suppression of HCT116 cells, suggesting that GRK5 mediates cell growth suppression by TIG1A. Thus, TIG1 may participate in the downregulation of G-protein coupled signaling by upregulating GRK5 expression.

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“…Real-time RT-PCR was performed to determine changes in FucT-VII gene expression as described by Miyashiro et al (2004), with some modification. First, HT-29 cells were pretreated by different LMW-ABP samples (5, 10, and 20 mg/mL) for 48 h. Second, total cellular RNA was isolated from HT-29 cells using an Trizol reagent kit (Invitrogen, MD) according to the supplier's instruction manuals.…”

Section: Analysis Of Fuct-vii Mrna Expression By Real-time Rt-pcrmentioning

confidence: 99%

“…Cell-surface antigen sLe x [NeuAca2-3Galb1-4 (Fuca1-3) GlcNAc-R] is a constituent of the ligand for cell adhesion molecules, particularly expressed on the surface of all circulating leukocytes or carcinoma cells. The biosynthesis of sLe x is mediated by FucT-VII, which catalyzes the transfer of fucose from GDP-b-fucose to the 3-OH of a-2,3-sialyl N-acetyllactosamine (SALN) (Miyashiro, Furuya, Fujishige, & Sugita, 2004;Zerfaoui, Fukuda, Sbarra, Lombardo, & EI-Battari, 2000). Many researchers believe that selectins, particularly E-selectin, are engaged in the growth of tumor and formation of metastasis, most probably through increased tumor angiogenesis and adhesion of tumor cells to endothelium (Alexandrakis, Passan, & Sfiridaki, 2004;Koch, Halloran, Haskell, Shah, & Polverini, 1995;Ley, 2003).…”

Section: Introductionmentioning

confidence: 99%

A polysaccharide isolated from Agaricus blazei Murill inhibits sialyl Lewis X/E-selectin-mediated metastatic potential in HT-29 cells through down-regulating α-1,3-fucosyltransferase-VII (FucT-VII)

Liu

Yue

Zhang

et al. 2010

Carbohydrate Polymers

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Highly sensitive cell-based assay system to monitor the sialyl Lewis X biosynthesis mediated by α1-3 fucosyltransferase-VII

Cited by 8 publications

References 37 publications

Variable DNA Methylation Is Associated with Chronic Obstructive Pulmonary Disease and Lung Function

Variable DNA Methylation Is Associated with Chronic Obstructive Pulmonary Disease and Lung Function

G protein-coupled receptor kinase 5 mediates Tazarotene-induced gene 1-induced growth suppression of human colon cancer cells

A polysaccharide isolated from Agaricus blazei Murill inhibits sialyl Lewis X/E-selectin-mediated metastatic potential in HT-29 cells through down-regulating α-1,3-fucosyltransferase-VII (FucT-VII)

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