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HNF4α is a Crucial Modulator of the Cholesterol-Dependent Regulation of NPC1L1

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Abstract

Purpose

Niemann-Pick C1-like 1 (NPC1L1) has been identified as a target of ezetimibe and found to be responsible for intestinal cholesterol absorption. Although, it was recently demonstrated that sterol responsive element binding protein 2 (SREBP2) is responsible for the cholesterol-dependent down-regulation of NPC1L1, the molecular mechanism of NPC1L1 expression is not fully understood. In the present study, we examined the involvement of hepatocyte nuclear factor 4α (HNF4α), a key modulator of lipid metabolism, in the transcriptional regulation of human NPC1L1 gene.

Methods

Reporter gene assays and EMSAs were performed using human NPC1L1 promoter constructs and the effect of siHNF4α was examined.

Results

Transfection of SREBP2 induced the transcriptional activities of NPC1L1 and additional transfection of HNF4α results in a marked stimulation of the activities. Studies with deletion mutants indicated that important elements are located within 264 nt upstream in the human NPC1L1 promoter. In addition, studies with mutations in putative binding sites of HNF4α indicated the existence of binding sites in −209 to −197 and −52 to −40. Moreover, HNF4α knockdown resulted in the reduced expression and regulation by cholesterol.

Conclusions

It is concluded that HNF4α plays a crucial role in the expression and regulation of human NPC1L1 gene.

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Abbreviations

DMEM:

Dulbecco’s Modified Eagle Medium

DR:

direct repeat

EMSA:

electrophoretic mobility shift assay

HMG-CoA:

3-hydroxy-3-methyl-glutaryl coenzyme A

HNF4α:

hepatocyte nuclear factor 4α

LDLR:

low density lipoprotein receptor

LXR:

liver X receptor

NF-Y:

nuclear factor-Y

NPC1L1:

Niemann-Pick C1-like 1

25-HCH:

25-hydroxy cholesterol

PGC-1α:

peroxisome proliferator-activated receptor (PPAR)-γ coactivator-1α

SRE:

sterol responsive element

SREBP:

sterol responsive element binding protein

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Acknowledgements

This work was supported by grants from The Japanese Ministry of Education, Science, Sports and Culture.

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Authors and Affiliations

  1. Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Yuki Iwayanagi, Tappei Takada & Hiroshi Suzuki

Authors
  1. Yuki Iwayanagi
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  2. Tappei Takada
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  3. Hiroshi Suzuki
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Correspondence to Tappei Takada.

Additional information

Yuki Iwayanagi and Tappei Takada are equally contributed.

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Iwayanagi, Y., Takada, T. & Suzuki, H. HNF4α is a Crucial Modulator of the Cholesterol-Dependent Regulation of NPC1L1. Pharm Res 25, 1134–1141 (2008). https://doi.org/10.1007/s11095-007-9496-9

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  • DOI: https://doi.org/10.1007/s11095-007-9496-9

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