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J. Lipid Res.
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The Journal of Lipid Research, Vol. 39, 2048-2053, October 1998
Copyright © 1998 by Lipid Research, Inc.

A novel 3T3-L1 preadipocyte variant that expresses PPAR{gamma}2 and RXR{alpha} but does not undergo differentiation

Rebecca A. Bailliea, Xiaoming Shaa, Philippe Thuilliera, and Steven D. Clarkea
a Institute for Cellular and Molecular Biology, and the Division of Nutritional Sciences, The University of Texas, Austin, Texas 78712

Correspondence to: Steven D. Clarke.

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 foci, 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 {gamma}2 (PPAR{gamma}2) or its heterodimer partner, retinoid X receptor {alpha} (RXR{alpha}). Surprisingly, Western blots revealed that 3T3-L1/RB1 cells contained both PPAR{gamma}2 and RXR{alpha} 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 PPAR{gamma}2 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 PPAR{gamma}2, 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 PPAR{gamma}2 and RXR{alpha} but does not undergo differentiation. J. Lipid Res. 1998. 39: 2048–2053.

Supplementary key words: fatty acid binding protein, adipocytes, differentiation, 3T3-L1 cells, PPAR


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