Yonsei Med J.  2006 Oct;47(5):729-736. 10.3349/ymj.2006.47.5.729.

Inflammatory Gene Expression Patterns Revealed by DNA Microarray Analysis in TNF-alpha-treated SGBS Human Adipocytes

• Do MS
• Jeong HS
• Choi BH
• Hunter L
• Langley S
• Pazmany L
• Trayhurn P

• KMID: 1381255
• DOI: http://doi.org/10.3349/ymj.2006.47.5.729

Abstract

We report here the use of human inflammation arrays to study the inflammatory gene expression profile of TNF-alpha- treated human SGBS adipocytes. Human preadipocytes (SGBS) were induced to differentiate in primary culture, and adipocyte differentiation was confirmed, using Oil Red O staining. We treated the differentiated adipocytes with TNF-alpha, and RNA from differentiated adipocytes with or without TNF-alpha treatment was hybridized to MWG human inflammation arrays to compare expression profiles. Eleven genes were up- or down-regulated in TNF-alpha-treated adipocytes. As revealed by array analysis, among 6 up-regulated genes, only eotaxin-1, monocyte chemoattractant protein-1 (MCP-1), and vascular cell adhesion molecule 1 isoform a precursor (VCAM1) were confirmed by real-time polymerase chain reaction (PCR). Similarly, among 5 down-regulated genes, only IL-1 family member 5 (IL1F5), a disintegrin and metalloprotease with thrombospondin motifs-1 preproprotein (ADAMTS1), fibronectin 1 isoform 1 preprotein (FN1), and matrix metalloproteinase 15 preprotein (MMP15) were confirmed by real-time PCR. There was a substantial increase (50-fold) in eotaxin-1 in response to TNF-alpha. Taken together, we have identified several inflammatory molecules expressed in SGBS adipocytes and discovered molecular factors explaining the relationship between obesity and atherosclerosis, focusing on inflammatory cytokines expressed in the TNF-alpha-treated SGBS cells. Further investigation into the role of these up- or down-regulated cytokine genes during the pathological processes leading to the development of atherosclerosis is warranted.

Keyword

Adipose tissue; cytokine; DNA microarray; inflammation; SGBS; TNF-alpha

MeSH Terms

Tumor Necrosis Factor-alpha/*pharmacology
Polymerase Chain Reaction
Oligonucleotide Array Sequence Analysis
Inflammation Mediators/*metabolism
Humans
Gene Expression Regulation/*drug effects
Gene Expression Profiling
Cells, Cultured
Cell Differentiation
Adipocytes/cytology/drug effects/*metabolism

Figure

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  Fig. 1 Oil Red O staining of SGBS cells at (A) Day 0 and at (B) Day 4, (C) Day 10, and (D) Day 20 after differentiation. The stained lipid droplets are shown in red.

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  Fig. 2 Scanned image data of a MWG human inflammatory array, hybridized with cRNA prepared from differentiated SGBS (Day 20) and TNF-α-treated differentiated SGBS cells (Day 20/TNF-α). (A) Day 20 cells are Cy3-labeled and Day 20/TNF-α cells are Cy5-labeled. (B) Day 20 cells are Cy5-labeled and Day 20/TNF-α cells are Cy3-labeled.

Reference

1. Kershaw EE, Flier JS. Adipose tissue as an endocrine organ. J Clin Endocrinol Metab. 2004. 89:2548–2556.

2. Trayhurn P, Wood IS. Signalling role of adipose tissue: adipokines and inflammation in obesity. Biochem Soc Trans. 2005. 33:1078–1081.

3. Fantuzzi G. Adipose tissue, adipokines, and inflammation. J Allergy Clin Immunol. 2005. 15:911–920.

4. Ziccardi P, Nappo F, Giugliano G, Esposito K, Marfella R, Cioffi M, et al. Reduction of inflammatory cytokine concentrations and improvement of endothelial functions in obese women after weight loss over one year. Circulation. 2002. 105:804–809.

5. Yudkin JS. Adipose tissue, insulin action and vascular disease: inflammatory signals. Int J Obes Relat Metab Disord. 2003. 27:Suppl 3. S25–S28.

6. Sethi JK, Hotamisligil GS. The role of TNFα in adipocyte metabolism. Semin Cell Dev Biol. 1999. 10:19–29.

7. Grunfeld C, Feingold KR. Regulation of lipid metabolism by cytokines during host defense. Nutrition. 1996. 12(1):Suppl. S24–S26.

8. Wabitsch M, Brenner RE, Melzner I, Braun M, Moller P, Heinze E, et al. Characterization of a human preadipocyte cell strain with high capacity for adipose differentiation. Int J Obes Relat Metab Disord. 2001. 25:8–15.

9. Sartipy P, Loskutoff DJ. Monocyte chemoattractant protein 1 in obesity and insulin resistance. Proc Natl Acad Sci USA. 2003. 100:7265–7270.

10. Takahashi K, Mizuarai S, Araki H, Mashiko S, Ishihara A, Kanatani A, et al. Adiposity elevates plasma MCP-1 levels leading to the increased CD11b-positive monocytes in mice. J Biol Chem. 2003. 278:46654–46660.

11. Kodali RB, Kim WJ, Galaria II, Miller C, Schecter AD, Lira SA, et al. CCL11 (Eotaxin) induces CCR3-dependent smooth muscle cell migration. Arterioscler Thromb Vasc Biol. 2004. 24:1211–1216.

12. De Caterina R, Bernini W, Carluccio MA, Liao JK, Libby P. Structural requirements for inhibition of cytokine-induced endothelial activation by unsaturated fatty acids. J Lipid Res. 1998. 39:1062–1070.

13. Lyon CJ, Law RE, Hsueh WA. Minireview: adiposity, inflammation, and atherogenesis. Endocrinology. 2003. 144:2195–2200.

14. Stetler-Stevenson WG, Liotta LA, Kleiner DE Jr. Extracellular matrix 6: role of matrix metalloproteinases in tumor invasion and metastasis. FASEB J. 1993. 7:1434–1441.

15. Hynes RO. The dynamic dialogue between cells and matrices: implications of fibronectin's elasticity. Proc Natl Acad Sci USA. 1999. 96:2588–2590.

16. Caterson B, Flannery CR, Hughes CE, Little CB. Mechanisms involved in cartilage proteoglycan catabolism. Matrix Biol. 2000. 19:333–344.

17. Stanton H, Ung L, Fosang AJ. The 45 kDa collagen-binding fragment of fibronectin induces matrix metalloproteinase-13 synthesis by chondrocytes and aggrecan degradation by aggrecanases. Biochem J. 2002. 364:181–190.

18. Okada Y. Ruddy S, Harris ED, Sledge CB, editors. Proteinase and matrix degradation. Kelly's Textbook of Rheumatology. 2001. Philadelphia: WB Saunders Company;55–72.

19. Tetlow LC, Adlam DJ, Woolley DE. Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage: associations with degenerative changes. Arthritis Rheum. 2001. 44:585–594.

20. Voros G, Maquoi E, Collen D, Lijnen HR. Differential expression of plasminogen activator inhibitor-1, tumor necrosis factor-alpha, TNF-alpha converting enzyme and ADAMTS family members in murine fat territories. Biochim Biophys Acta. 2003. 1625:36–42.

21. Boeuf S, Klingenspor M, Van Hal NL, Schneider T, Keijer J, Klaus S. Differential gene expression in white and brown preadipocytes. Physiol Genomics. 2001. 7:15–25.

22. Debets R, Timans JC, Homey B, Zurawski S, Sana TR, Lo S, et al. Two novel IL-1 family members, IL-1delta and IL-1epsilon function as an antagonist and agonist of NF-κB activation through the orphan IL-1 receptor-related protein 2. J Immunol. 2001. 167:1440–1446.