A novel 3T3-L1 preadipocyte variant that expresses PPARγ2 and RXRα but does not undergo differentiation

Rebecca A. Baillie, Xiaoming Sha, Philippe Thuillier, Steven D. Clarke

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

This report describes a novel adipocyte-like cell line termed 3T3- L1/RB1 that was derived from preadipocyte cell line, 3T3-L1. The 3T3-L1/RB1 cells continued to divide after reaching confluence, formed loci, and constitutively expressed a low level of adipose fatty acid binding protein (A-FABP) mRNA. However, 3T3L-1/RB cells did not undergo terminal differentiation as indicated by the failure of insulin and thiazolidendiones to induce the expression of A-FABP, lipoprotein lipase, and fatty acid synthase. We hypothesized that the 3T3-L1/RB1 variant did not respond to differentiation stimuli because it did not express either peroxisomal proliferator activated receptor γ2 (PPARγ2) or its heterodimer partner, retinoid X receptor α (RXRα). Surprisingly, Western blots revealed that 3T3-L1/RB1 cells contained both PPARγ2 and RXRα proteins at levels equal to or greater than that of the parent cell line. However, gel retardation assays using the adipose response element from A-FABP and nuclear protein extracts from 3T3-L1/RB1 cells treated with insulin or pioglitazone revealed that nuclear protein extracts from 3T3-L1/RB1 cells had very little ability to bind the PPARγ2 recognition sequence of the A-FABP gene. These data suggest that the 3T3-L1/RB1 variant contains a mutation that may prevent ligand activation of PPARγ2, and the subsequent conversion of 3T3-L1/RB1 cells to mature fat cells. - Baillie, R. A., X. Sha, P. Thuillier, and S. D. Clarke. A novel 3T3-L1 preadipocyte variant that expresses PPARγ2 and RXRα but does not undergo differentiation.

Original languageEnglish (US)
Pages (from-to)2048-2053
Number of pages6
JournalJournal of Lipid Research
Volume39
Issue number10
StatePublished - Oct 1998
Externally publishedYes

Fingerprint

3T3-L1 Cells
Retinoid X Receptors
Fatty Acid-Binding Proteins
pioglitazone
Cells
Nuclear Proteins
Adipocytes
Cell Line
Insulin
Fatty Acid Synthases
Lipoprotein Lipase
Response Elements
Electrophoretic Mobility Shift Assay
Assays
Genes
Gels
Chemical activation
Fats
Ligands
Messenger RNA

Keywords

  • 3T3-L1 cells
  • Adipocytes
  • Differentiation
  • Fatty acid binding protein
  • PPAR

ASJC Scopus subject areas

  • Endocrinology

Cite this

A novel 3T3-L1 preadipocyte variant that expresses PPARγ2 and RXRα but does not undergo differentiation. / Baillie, Rebecca A.; Sha, Xiaoming; Thuillier, Philippe; Clarke, Steven D.

In: Journal of Lipid Research, Vol. 39, No. 10, 10.1998, p. 2048-2053.

Research output: Contribution to journalArticle

Baillie, Rebecca A. ; Sha, Xiaoming ; Thuillier, Philippe ; Clarke, Steven D. / A novel 3T3-L1 preadipocyte variant that expresses PPARγ2 and RXRα but does not undergo differentiation. In: Journal of Lipid Research. 1998 ; Vol. 39, No. 10. pp. 2048-2053.
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