Regulation of human β-galactoside α2,6-sialyltransferase (hST6Gal I) gene expression during differentiation of human osteoblastic MG-63 cells

An, So-Young; Lee, Miri; Yoon, Hye‐Jung; Abekura, Fukushi; Kim, Kyoung-Sook; Kim, Dong-Hyun; Kim, Hyeon-Jun; Lee, Kichoon; Kim, Cheorl-Ho; Lee, Young Choon

doi:10.1007/s10719-020-09959-3

Cited by 6 publications

(18 citation statements)

References 29 publications

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“…Total RNA was extracted from cells treated with various concentrations of curcumin for 24 h using Trizol reagent (Invitrogen). RT-PCR was performed using gene-specific primers ( Table 1 ) as previously described ( Lee et al, 2018a , 2018b ; An et al, 2020 ). PCR products were analyzed by 1.5% agarose gel electrophoresis and visualized with ethidium bromide.…”

Section: Methodsmentioning

confidence: 99%

“…We isolated the 5′-flanking region of the hST6GalNAc I gene by PCR using the sequence information of the National Center for Biotechnology Information (NCBI Reference Sequence NC_000017.11). This was conducted in a similar manner to the experimental method described previously ( An et al, 2020 ). A 1974 bp fragment of the 5′-flanking sequence of the hST6GalNAc I gene were amplified by long and accurate PCR (LA-PCR) using human genomic DNA (Clontech) as template, and a sense primer P-1974S and an antisense primer P + 1A containing Kpn I and Xho I sites, respectively ( Table 1 ).…”

Section: Methodsmentioning

confidence: 99%

“…ChIP assay was conducted to assess the interaction between ERα and ERE located in hST6GalNAc I promoter using the ChIP kit (Upstate Biotechonology) according to the same protocol described previously ( Lee et al, 2018a , 2018b ; An et al, 2020 ). Immunoprecipitation was performed using 5 µg of anti-ERα antibody (ab32063, Abcam) and IgG antibodies (Sigma).…”

Section: Methodsmentioning

confidence: 99%

“…ChIP assay was conducted to assess the interaction between ERα and ERE located in hST6GalNAc I promoter using the ChIP kit (Upstate Biotechonology) according to the same protocol described previously (Lee et al, 2018a(Lee et al, , 2018bAn et al, 2020).…”

Section: Chromatin Immunoprecipitation Assaymentioning

confidence: 99%

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Curcumin-mediated transcriptional regulation of human N-acetylgalactosamine-α2,6-sialyltransferase which synthesizes sialyl-Tn antigen in HCT116 human colon cancer cells

Kim

Cho

et al. 2022

Front. Mol. Biosci.

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Human N-acetylgalactosamine-α2,6-sialyltransferase (hST6GalNAc I) is the major enzyme involved in the biosynthesis of sialyl-Tn antigen (sTn), which is known to be expressed in more than 80% of human carcinomas and correlated with poor prognosis in cancer patients. Athough high expression of hST6GalNAc I is associated with augmented proliferation, migration and invasion in various cancer cells, transcriptional mechanism regulating hST6GalNAc I gene expression remains largely unknown. In this study, we found that hST6GalNAc I gene expression was markedly augmented by curcumin in HCT116 human colon carcinoma cells. To understand the molecular mechanism for the upregulation of hST6GalNAc I gene expression by curcumin in HCT116 cells, we first determined the transcriptional start site of hST6GalNAc I gene by 5′-RACE and cloned the proximal hST6GalNAc I 5′-flanking region spanning about 2 kb by PCR. Functional analysis of the hST6GalNAc I 5′ flanking region of hST6GalNAc I by sequential 5′-deletion, transient transfection of reporter gene constructs and luciferase reporter assays showed that -378/-136 region is essential for maximal activation of transcription in response to curcumin in HCT 116 cells. This region includes putative binding sites for transcription factors c-Ets-1, NF-1, GATA-1, ER-α, YY1, and GR-α. ChIP analysis and site-directed mutagenesis demonstrated that estrogen receptor α (ER-α) binding site (nucleotides -248/-238) in this region is crucial for hST6GalNAc I gene transcription in response to curcumin stimulation in HCT116 cells. The transcription activity of hST6GalNAc I gene induced by curcumin in HCT116 cells was strongly inhibited by PKC inhibitor (Gö6983) and ERK inhibitor (U0126). These results suggest that curcumin-induced hST6GalNAc I gene expression in HCT116 cells is modulated through PKC/ERKs signal pathway.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…ChIP assay was conducted to assess the interaction between ERα and ERE located in hST6GalNAc I promoter using the ChIP kit (Upstate Biotechonology) according to the same protocol described previously (Lee et al, 2018a(Lee et al, , 2018bAn et al, 2020).…”

Section: Chromatin Immunoprecipitation Assaymentioning

confidence: 99%

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Curcumin-mediated transcriptional regulation of human N-acetylgalactosamine-α2,6-sialyltransferase which synthesizes sialyl-Tn antigen in HCT116 human colon cancer cells

Kim

Cho

et al. 2022

Front. Mol. Biosci.

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“…When the cells were further cultured in the RPMI-1640 complete medium, lipid raftspecific Sia was rapidly reconditioned and almost completely renewed to 97.1% within 24 h. Interestingly, cellular lipid raftspecific Sia was over-repaired to 127.1% during continued incubation from 24 to 36 h. This may be attributable to an increase in sialyltransferase levels that was not decreased in a timely manner. 49 When the cells were further incubated for up to 48 h, we found that lipid raft-specific Sia levels returned to normal after excessive repair (Figure 3). In contrast to the dynamic and constant renewal of raft-specific Sia, the signaling of raft-specific proteins remained stable, indicating that changes in Sia did not affect the binding of HRP-CTxB (Figure 3).…”

Section: ■ Results and Discussionmentioning

confidence: 94%

Multifunctional Proximity Labeling Strategy for Lipid Raft-Specific Sialic Acid Tracking and Engineering

Guo,

Wang,

Jiang

et al. 2023

Bioconjugate Chem.

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Lipid raft-specific glycosylation has been implicated in many biological processes, including intracellular trafficking, cell adhesion, signal transduction, and host–pathogen interactions. The major predicament in lipid raft-specific glycosylation research is the unavailability of tools for tracking and manipulating glycans on lipid rafts at the microstructural level. To overcome this challenge, we developed a multifunctional proximity labeling (MPL) platform that relies on cholera toxin B subunit to localize horseradish peroxidase on lipid rafts. In addition to the prevailing electron-rich amino acids, modified sialic acid was included in the horseradish peroxidase-mediated proximity labeling substrate via purposefully designed chemical transformation reactions. In combination with sialic acid editing, the self-renewal of lipid raft-specific sialic acid was visualized. The MPL method enabled tracking of lipid raft dynamics under methyl-β-cyclodextrin and mevinolin treatments; in particular, the alteration of lipid rafts markedly affected cell migration. Furthermore, we embedded functional molecules into the method and implemented raft-specific sialic acid gradient engineering. Our novel strategy presents opportunities for tailoring lipid raft-specific sialic acids, thereby regulating interactions associated with lipid raft regions (such as cell–virus and cell–microenvironment interactions), and can aid in the development of lipid raft-based therapeutic regimens for tumors.

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Regulatory mechanism for the human glioblastoma cell-specific expression of the human GD1c/GT1a/GQ1b synthase (hST8Sia V) gene

An,

Lee,

Kim

et al. 2023

Glycoconj J

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In this study we observed that human GD1c/GT1a/GQ1b synthase (hST8Sia V) is particularly expressed in human glioblastoma cells. To address the mechanism regulating human glioblastoma-speci c gene expression of the hST8Sia V, after the transcription start site (TSS) was identi ed by the 5'-rapid ampli cation of cDNA end with total RNA from human glioblastoma U87MG cells, the 5'-anking region (2.5 kb) of the hST8Sia V gene was isolated and its promoter activity was examined. By luciferase reporter assay, this 5'-anking region revealed strong promoter activity in only U-87MG cells, but not in other tissue-derived cancer cells. 5'-deletion mutant analysis showed that the region from -1140 to -494 is crucial for transcription of the hST8Sia V gene in U87MG cells. This region contains the activator protein-1 (AP-1) binding site, the main target of the c-Jun N-terminal kinase (JNK) downstream. The AP-1 binding site at -1043/-1037 was proved to be indispensable for the hST8Sia V gene-speci c expression in U87MG cells by site-directed mutagenesis. Moreover, the transcriptional activation of hST8Sia V gene in U87MG cells was strongly inhibited by a speci c JNK inhibitor, SP600125. These results suggest that the hST8Sia V gene-speci c expression in U87MG cells is controlled by JNK/AP-1 signaling pathway.

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Regulation of human β-galactoside α2,6-sialyltransferase (hST6Gal I) gene expression during differentiation of human osteoblastic MG-63 cells

Cited by 6 publications

References 29 publications

Curcumin-mediated transcriptional regulation of human N-acetylgalactosamine-α2,6-sialyltransferase which synthesizes sialyl-Tn antigen in HCT116 human colon cancer cells

Curcumin-mediated transcriptional regulation of human N-acetylgalactosamine-α2,6-sialyltransferase which synthesizes sialyl-Tn antigen in HCT116 human colon cancer cells

Multifunctional Proximity Labeling Strategy for Lipid Raft-Specific Sialic Acid Tracking and Engineering

Regulatory mechanism for the human glioblastoma cell-specific expression of the human GD1c/GT1a/GQ1b synthase (hST8Sia V) gene

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