Suppression of Transcription Factor PDX-1/IPF1/STF-1/IDX-1 Causes No Decrease in Insulin mRNA in MIN6 Cells

Kajimoto, Yoshitaka; Watada, Hirotaka; Matsuoka, Taka-aki; Kaneto, Hideaki; Fujitani, Yoshio; Miyazaki, Jun-ichi; Yamasaki, Yoshimitsu

doi:info:doi/10.1172/JCI119712

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タイトル	Suppression of Transcription Factor PDX-1/IPF1/STF-1/IDX-1 Causes No Decrease in Insulin mRNA in MIN6 Cells
別タイトル	nichi , Yamasaki, Yoshimitsu
著者	Kajimoto, Yoshitaka Kajimoto, Yoshitaka
	Watada, Hirotaka Watada, Hirotaka
	Matsuoka, Taka-aki Matsuoka, Taka-aki
	Kaneto, Hideaki Kaneto, Hideaki
	Fujitani, Yoshio Fujitani, Yoshio
	Miyazaki, Jun-ichi Miyazaki, Jun-ichi
	Yamasaki, Yoshimitsu Yamasaki, Yoshimitsu
キーワード等	gene expression regulation
	transcription factors
	homeodomain protein
	antisense oligodeoxynucleotide
抄録	The insulin gene transcription factor PDX-1/IPF1/STF-1/ IDX-1 plays a key role in directing beta cell-specific gene expressions. Recently, impairment of PDX-1 expression or activity has been observed in beta cell-derived HIT cells cultured under high glucose concentrations, and this has been suggested as a possible cause of the decrease in insulin gene transcription. To investigate the pathophysiological significance of PDX-1 as a determinant of the rate of insulin gene transcription, we suppressed its expression in beta cell-derived MIN6 cells using an antisense oligodeoxynucleotide (ODN) and searched for possible changes in the beta cell-specific gene expression. Treatment of MIN6 cells with an 18-mer phosphorothioate ODN complementary to a sequence starting at the translation initiation codon of PDX-1 caused a potent, concentration-dependent reduction in PDX-1 expression; addition of 2 microM antisense ODN could reduce PDX-1 expression to 14+/-4% of the control. There was also a decrease in its DNA binding to the insulin gene A element. Despite such suppression of PDX-1, Northern blot analysis revealed no decrease in the amount of insulin mRNA in the MIN6 cells. Similarly, no changes were detected in the transcription of the glucokinase or islet amyloid polypeptide gene, for which PDX-1 was shown to function as a transcription factor. Thus, our findings dispute the physiological significance of PDX-1 in determining the rate of insulin gene transcription. This means that other components constituting the transcription-controlling machinery need to be evaluated in order to understand the molecular basis of impaired insulin biosynthesis such as that observed due to glucose toxicity.
公開者	American Society for Clinical Investigation
掲載誌名	Journal of Clinical Investigation
巻	100
号	7
開始ページ	1840
終了ページ	1846
刊行年月	1997-10
ISSN	00219738
NCID	AA00695520
DOI	info:doi/10.1172/JCI119712
URL	http://hdl.handle.net/11094/23107
権利情報	Copyright © 1997 The American Society for Clinical Investigation
言語	英語
カテゴリ	学術雑誌論文 Journal Article

著者版フラグ	publisher
NII資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
dcmi資源タイプ	text
DCTERMS.bibliographicCitation	Journal of Clinical Investigation.100(7) P.1840-P.1846
DC.title	Suppression of Transcription Factor PDX-1/IPF1/STF-1/IDX-1 Causes No Decrease in Insulin mRNA in MIN6 Cells
DCTERMS.alternative	nichi , Yamasaki, Yoshimitsu
DC.creator	Kajimoto, Yoshitaka
	Watada, Hirotaka
	Matsuoka, Taka-aki
	Kaneto, Hideaki
	Fujitani, Yoshio
	Miyazaki, Jun-ichi
	Yamasaki, Yoshimitsu
DC.publisher	American Society for Clinical Investigation
DC.language" scheme="DCTERMS.RFC1766	英語
DCTERMS.issued" scheme="DCTERMS.W3CDTF	1997-10
DC.identifier	info:doi/10.1172/JCI119712
DC.identifier" scheme="DCTERMS.URI	http://hdl.handle.net/11094/23107
DC.subject	gene expression regulation
	transcription factors
	homeodomain protein
	antisense oligodeoxynucleotide
DCTERMS.abstract	The insulin gene transcription factor PDX-1/IPF1/STF-1/ IDX-1 plays a key role in directing beta cell-specific gene expressions. Recently, impairment of PDX-1 expression or activity has been observed in beta cell-derived HIT cells cultured under high glucose concentrations, and this has been suggested as a possible cause of the decrease in insulin gene transcription. To investigate the pathophysiological significance of PDX-1 as a determinant of the rate of insulin gene transcription, we suppressed its expression in beta cell-derived MIN6 cells using an antisense oligodeoxynucleotide (ODN) and searched for possible changes in the beta cell-specific gene expression. Treatment of MIN6 cells with an 18-mer phosphorothioate ODN complementary to a sequence starting at the translation initiation codon of PDX-1 caused a potent, concentration-dependent reduction in PDX-1 expression; addition of 2 microM antisense ODN could reduce PDX-1 expression to 14+/-4% of the control. There was also a decrease in its DNA binding to the insulin gene A element. Despite such suppression of PDX-1, Northern blot analysis revealed no decrease in the amount of insulin mRNA in the MIN6 cells. Similarly, no changes were detected in the transcription of the glucokinase or islet amyloid polypeptide gene, for which PDX-1 was shown to function as a transcription factor. Thus, our findings dispute the physiological significance of PDX-1 in determining the rate of insulin gene transcription. This means that other components constituting the transcription-controlling machinery need to be evaluated in order to understand the molecular basis of impaired insulin biosynthesis such as that observed due to glucose toxicity.
DC.rights	Copyright © 1997 The American Society for Clinical Investigation
citation_title	Suppression of Transcription Factor PDX-1/IPF1/STF-1/IDX-1 Causes No Decrease in Insulin mRNA in MIN6 Cells
citation_author	Kajimoto, Yoshitaka
	Watada, Hirotaka
	Matsuoka, Taka-aki
	Kaneto, Hideaki
	Fujitani, Yoshio
	Miyazaki, Jun-ichi
	Yamasaki, Yoshimitsu
citation_publisher	American Society for Clinical Investigation
citation_language	英語
citation_date	1997-10
citation_journal_title	Journal of Clinical Investigation
citation_volume	100
citation_issue	7
citation_firstpage	1840
citation_lastpage	1846
citation_issn	00219738
citation_doi	info:doi/10.1172/JCI119712
citation_public_url	http://hdl.handle.net/11094/23107
citation_keywords	gene expression regulation
	transcription factors
	homeodomain protein
	antisense oligodeoxynucleotide

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