Enhancive effects of d-glucose and its analogs on expression of d-glucose-unrelated transgenes in mammalian cells

Miyuki Kimura, Hikaru Nanba, Manabu Okubo, Mai Ezumi, Nao Susumu, Masao Yamada, Yujiro Arao

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We studied the effects of d-glucose on transgene expression in mammalian cells by a reporter gene assay using CV-1 cells and a CMV promoter-controlled EGFP gene. Treatment of CV-1 cells with 5% d-glucose unchanged the number of fluorescent cells in fluorescence microscopic observation but significantly intensified fluorescence in the fluorometric assay. Furthermore, EGFP itself and mRNA became more abundant in Western blot and quantitative RT-PCR analyses of 5% d-glucose-treated cells, respectively. These results indicate that elevated d-glucose can activate transgene expression via transcriptional stimulation, at least in part. The same concentrations of l-glucose led to only negligible increases in transgene expression, indicating that d-glucose's effect is different from its osmotic effect. The d-glucose-induced augmentation of fluorescence was observed not only in the experiment using the CMV promoter-controlled EGFP gene but also in experiments using the SV40 and RSV promoter-controlled ones, suggesting that elevated d-glucose can enhance transgene expression regulated by various promoters commonly used in transgene expression. The assessment of d-glucose analogs for their enhancive effects on transgene expression revealed that 1,6-anhydro- d-glucose and β-methyl- d-glucoside had stronger effects than d-glucose. From this result, we can expect to find more effective carbohydrates to enhance transgene expression. The α- and β-M- d-glucosides, which are slightly different from each other in three-dimensional structure, exerted largely distinct stimulative effects on transgene expression, suggesting that fundamental rules determine the enhancive effects of saccharides and that the modification of the saccharide by applying such rules will enable us to develop more powerful substances for transgene expression.

Original languageEnglish
Pages (from-to)194-201
Number of pages8
JournalJournal of Bioscience and Bioengineering
Volume112
Issue number2
DOIs
Publication statusPublished - Aug 2011

Fingerprint

Transgenes
Glucose
Cells
Genes
Fluorescence
Glucosides
Assays
Carbohydrates
Reporter Genes
Cell Count
Western Blotting
Experiments
Polymerase Chain Reaction
Messenger RNA

Keywords

  • D-Glucose
  • Enhancement
  • Monosaccharides
  • Transgene expression
  • Virus promoter

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Enhancive effects of d-glucose and its analogs on expression of d-glucose-unrelated transgenes in mammalian cells. / Kimura, Miyuki; Nanba, Hikaru; Okubo, Manabu; Ezumi, Mai; Susumu, Nao; Yamada, Masao; Arao, Yujiro.

In: Journal of Bioscience and Bioengineering, Vol. 112, No. 2, 08.2011, p. 194-201.

Research output: Contribution to journalArticle

@article{09b071f3a9604a6180ed79147f690c58,
title = "Enhancive effects of d-glucose and its analogs on expression of d-glucose-unrelated transgenes in mammalian cells",
abstract = "We studied the effects of d-glucose on transgene expression in mammalian cells by a reporter gene assay using CV-1 cells and a CMV promoter-controlled EGFP gene. Treatment of CV-1 cells with 5{\%} d-glucose unchanged the number of fluorescent cells in fluorescence microscopic observation but significantly intensified fluorescence in the fluorometric assay. Furthermore, EGFP itself and mRNA became more abundant in Western blot and quantitative RT-PCR analyses of 5{\%} d-glucose-treated cells, respectively. These results indicate that elevated d-glucose can activate transgene expression via transcriptional stimulation, at least in part. The same concentrations of l-glucose led to only negligible increases in transgene expression, indicating that d-glucose's effect is different from its osmotic effect. The d-glucose-induced augmentation of fluorescence was observed not only in the experiment using the CMV promoter-controlled EGFP gene but also in experiments using the SV40 and RSV promoter-controlled ones, suggesting that elevated d-glucose can enhance transgene expression regulated by various promoters commonly used in transgene expression. The assessment of d-glucose analogs for their enhancive effects on transgene expression revealed that 1,6-anhydro- d-glucose and β-methyl- d-glucoside had stronger effects than d-glucose. From this result, we can expect to find more effective carbohydrates to enhance transgene expression. The α- and β-M- d-glucosides, which are slightly different from each other in three-dimensional structure, exerted largely distinct stimulative effects on transgene expression, suggesting that fundamental rules determine the enhancive effects of saccharides and that the modification of the saccharide by applying such rules will enable us to develop more powerful substances for transgene expression.",
keywords = "D-Glucose, Enhancement, Monosaccharides, Transgene expression, Virus promoter",
author = "Miyuki Kimura and Hikaru Nanba and Manabu Okubo and Mai Ezumi and Nao Susumu and Masao Yamada and Yujiro Arao",
year = "2011",
month = "8",
doi = "10.1016/j.jbiosc.2011.04.010",
language = "English",
volume = "112",
pages = "194--201",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "The Society for Biotechnology, Japan",
number = "2",

}

TY - JOUR

T1 - Enhancive effects of d-glucose and its analogs on expression of d-glucose-unrelated transgenes in mammalian cells

AU - Kimura, Miyuki

AU - Nanba, Hikaru

AU - Okubo, Manabu

AU - Ezumi, Mai

AU - Susumu, Nao

AU - Yamada, Masao

AU - Arao, Yujiro

PY - 2011/8

Y1 - 2011/8

N2 - We studied the effects of d-glucose on transgene expression in mammalian cells by a reporter gene assay using CV-1 cells and a CMV promoter-controlled EGFP gene. Treatment of CV-1 cells with 5% d-glucose unchanged the number of fluorescent cells in fluorescence microscopic observation but significantly intensified fluorescence in the fluorometric assay. Furthermore, EGFP itself and mRNA became more abundant in Western blot and quantitative RT-PCR analyses of 5% d-glucose-treated cells, respectively. These results indicate that elevated d-glucose can activate transgene expression via transcriptional stimulation, at least in part. The same concentrations of l-glucose led to only negligible increases in transgene expression, indicating that d-glucose's effect is different from its osmotic effect. The d-glucose-induced augmentation of fluorescence was observed not only in the experiment using the CMV promoter-controlled EGFP gene but also in experiments using the SV40 and RSV promoter-controlled ones, suggesting that elevated d-glucose can enhance transgene expression regulated by various promoters commonly used in transgene expression. The assessment of d-glucose analogs for their enhancive effects on transgene expression revealed that 1,6-anhydro- d-glucose and β-methyl- d-glucoside had stronger effects than d-glucose. From this result, we can expect to find more effective carbohydrates to enhance transgene expression. The α- and β-M- d-glucosides, which are slightly different from each other in three-dimensional structure, exerted largely distinct stimulative effects on transgene expression, suggesting that fundamental rules determine the enhancive effects of saccharides and that the modification of the saccharide by applying such rules will enable us to develop more powerful substances for transgene expression.

AB - We studied the effects of d-glucose on transgene expression in mammalian cells by a reporter gene assay using CV-1 cells and a CMV promoter-controlled EGFP gene. Treatment of CV-1 cells with 5% d-glucose unchanged the number of fluorescent cells in fluorescence microscopic observation but significantly intensified fluorescence in the fluorometric assay. Furthermore, EGFP itself and mRNA became more abundant in Western blot and quantitative RT-PCR analyses of 5% d-glucose-treated cells, respectively. These results indicate that elevated d-glucose can activate transgene expression via transcriptional stimulation, at least in part. The same concentrations of l-glucose led to only negligible increases in transgene expression, indicating that d-glucose's effect is different from its osmotic effect. The d-glucose-induced augmentation of fluorescence was observed not only in the experiment using the CMV promoter-controlled EGFP gene but also in experiments using the SV40 and RSV promoter-controlled ones, suggesting that elevated d-glucose can enhance transgene expression regulated by various promoters commonly used in transgene expression. The assessment of d-glucose analogs for their enhancive effects on transgene expression revealed that 1,6-anhydro- d-glucose and β-methyl- d-glucoside had stronger effects than d-glucose. From this result, we can expect to find more effective carbohydrates to enhance transgene expression. The α- and β-M- d-glucosides, which are slightly different from each other in three-dimensional structure, exerted largely distinct stimulative effects on transgene expression, suggesting that fundamental rules determine the enhancive effects of saccharides and that the modification of the saccharide by applying such rules will enable us to develop more powerful substances for transgene expression.

KW - D-Glucose

KW - Enhancement

KW - Monosaccharides

KW - Transgene expression

KW - Virus promoter

UR - http://www.scopus.com/inward/record.url?scp=79960710819&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79960710819&partnerID=8YFLogxK

U2 - 10.1016/j.jbiosc.2011.04.010

DO - 10.1016/j.jbiosc.2011.04.010

M3 - Article

C2 - 21596618

AN - SCOPUS:79960710819

VL - 112

SP - 194

EP - 201

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 2

ER -