Synthesis and secretion of apoC-I and apoE by human SW872 liposarcoma cells
ABSTRACT Apolipoprotein C-I (apoC-I) plays an important role in the metabolism of plasma triglyceride levels and cholesterol metabolism. Little is known about the regulation of apoC-I production by human adipocytes. Aim. To investigate the effect of different tissue culture conditions on the synthesis and secretion of apoC-I and apoE in human SW872 liposarcoma cells and to study the effects of apoC-I overexpression in these same cells. Methods. SW872 cells were grown in DMEM/F-12 (3:1, v/v). QPCR was used to quantify mRNA synthesis. ELISAs were used to quantify intracellular and extracellular proteins. Colorimetric reaction kits were used to analyze intracellular cholesterol and triglyceride concentrations. Results . Maturation experiments revealed that after 17 days in culture, SW872 cells contained significantly more cholesterol (100%) and triglyceride (3-fold). Cell maturation was associated with significantly higher levels of apoE mRNA (+200%) but not apoC-I mRNA (-50%). The cells secreted more apoC-I (+110%) and apoE (+340%). Cellular apoC-I increased 620% and apoE increased 1540%. Treatment of cells during maturation with insulin (0, 10 or 1000 nM) significantly reduced the secretion of apoC-I and apoE (-14% and -56%, respectively) and intracellular apoC-I and apoE (-10% and -12%, respectively. Overexpression of apoC-I in SW872 cells resulted in increased cell number (+70%) and decreased lipids per cell (-32% triglyceride, -36% cholesterol) as compared to controls. Conclusion. These results suggest that apoC-I and apoE production is differentially regulated at the transcriptional level in adipocytes and that apoC-I and apoE play a role in the maturation of human adipocytes and may have an important role in mediating or regulating cell lipid accumulation. As well, overexpression of apoC-I in SW872 cells impedes cellular lipid accumulation and stimulates cellular proliferation.
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ABSTRACT: Esters of phthalic acid (phthalates) are largely used in industrial plastics, medical devices, and pharmaceutical formulations. They are easily released from plastics into the environment and can be found in measurable levels in human fluids. Phthalates are agonists for peroxisome proliferator-activated receptors (PPARs), through which they regulate translocator protein (TSPO; 18 kDa) transcription in a tissue-specific manner. TSPO is a drug- and cholesterol-binding protein involved in mitochondrial respiration, steroid formation, and cell proliferation. TSPO has been shown to increase during differentiation and decrease during maturation in mouse adipocytes. The purpose of this study was to establish the effect of mono-(2-ethylhexyl) phthalate (MEHP) on the differentiation of human SW 872 preadipocyte cells, and examine the role of TSPO in the process. After 4 days of treatment with 10 µM MEHP, we observed changes in the transcription of acetyl-CoA carboxylase alpha, adenosine triphosphate citrate lyase, glucose transporters 1 and 4, and the S100 calcium binding protein B, all of which are markers of preadipocyte differentiation. These observed gene expression changes coincided with a decrease in cellular proliferation without affecting cellular triglyceride content. Taken together, these data suggest that MEHP exerts a differentiating effect on human preadipocytes. Interestingly, MEHP was able to temporarily increase TSPO mRNA levels through the PPAR-α and β/δ pathways. These results suggest that TSPO can be considered an important player in the differentiation process itself, or alternatively a factor whose presence is essential for adipocyte development.PLoS ONE 12/2011; 6(12):e28750. DOI:10.1371/journal.pone.0028750 · 3.53 Impact Factor
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ABSTRACT: The risk of cardiovascular disease is inversely correlated to level of plasma HDL-c. Moreover, reverse cholesterol transport (RCT) from peripheral tissues to the liver is the most widely accepted mechanism linked to the anti-atherosclerotic activity of HDL. The apolipoprotein A-I (apoA-I) and the ABC transporters play a key role in this process.Adipose tissue constitutes the body's largest pool of free cholesterol. The adipose cell could therefore be regarded as a key factor in cholesterol homeostasis. The present study investigates the capacity of primary cultures of mature human adipocytes to release cholesterol and explores the relationships between apoA-I, ABCA1, and apoE as well as the signaling pathways that could be potentially involved. We demonstrate that apoA-I induces a strong increase in cholesterol release and apoE secretion from adipocytes, whereas it has no transcriptional effect on ABCA1 or apoE genes. Furthermore, brefeldin A (BFA), an intracellular trafficking inhibitor, reduces basal cholesterol and apoE secretion, but does not modify induction by apoA-I. The use of statins also demonstrates that apoA-I stimulated cholesterol release is independent of HMG-CoA reductase activation. Our work highlights the fact that adipose tissue, and particularly adipocytes, may largely contribute to RCT via a mechanism specifically regulated within these cells. This further supports the argument that adipose tissue must be regarded as a major factor in the development of cardiovascular diseases, in particular atherosclerosis.Lipids in Health and Disease 07/2010; 9:75. DOI:10.1186/1476-511X-9-75 · 2.31 Impact Factor