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Using Caco-2 Cells to Study Lipid Transport by the Intestine
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Using Caco-2 Cells to Study Lipid Transport by the Intestine

Published on: August 20, 2015

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7keto-stigmasterol and 7keto-cholesterol induce differential proteome changes to intestinal epitelial (Caco-2) cells

J M Laparra1, A Alfonso-García2, A Alegría2

  • 1Institute of Food Engineering for Development, Politechnical University of Valencia, Avda. Cami de Vera s/n, 46022 Valencia, Spain.

Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association
|July 5, 2015
PubMed

Insights

Oxysterols like 7keto-cholesterol and 7keto-stigmasterol significantly alter intestinal cell proteomes, impacting mitochondrial function and immune responses. This proteomic analysis reveals distinct cellular effects, questioning oxysterol safety and regulatory potential.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Proteomics

Background:

  • Oxysterols are known to trigger inflammatory, immune cytotoxic, and apoptotic processes.
  • Their impact on cellular proteomes, particularly in intestinal epithelial cells, remains underexplored.

Purpose of the Study:

  • To conduct a comparative proteomic analysis of intestinal epithelial cells exposed to 7keto-cholesterol and 7keto-stigmasterol.
  • To investigate the distinct effects of these oxysterols on cellular pathways.

Main Methods:

  • Intestinal epithelial cell cultures were incubated with 7keto-cholesterol or 7keto-stigmasterol (60 μM/24 h).
  • Nano-liquid chromatography coupled with TripleTOF mass spectrometry (nLC-TripleTOF) was used for proteomic analysis.
  • Principal component analysis (PCA) was employed to analyze proteome patterns.

Main Results:

  • PCA clearly discriminated between proteome perturbations induced by 7keto-cholesterol and 7keto-stigmasterol.
  • Affected proteins are involved in mitochondrial function, lipid homeostasis, inflammation, immunity, and cell proliferation.
  • Key proteins showing differential expression include macrophage migration inhibitory factor, apolipoprotein E, Bcl-2-associated transcription factor, and cellular retinoic acid-binding protein.
  • 7keto-stigmasterol exposure specifically increased ubiquitin-conjugating enzyme E2 and mitochondrial superoxide dismutase.

Conclusions:

  • Distinct proteomic profiles arise from exposure to 7keto-cholesterol versus 7keto-stigmasterol in intestinal cells.
  • These findings highlight the need for further safety assessments and regulatory considerations for oxysterols.
  • Oxysterols may influence intestinal and immune cell differentiation, function, environmental response, and nutrient absorption.
Keywords:
7keto-cholesterol7keto-stigmasterolCaco-2 cellsOxysterolsProteome

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