Yonsei Medical Journal Vol. 47, No. 5, pp. 729 - 736, 2006 Inflammatory Gene Expression Patterns Revealed by DNA Microarray Analysis in TNF-α-treated SGBS Human Adipocytes Myoung-Sool Do,1 Hun-Soon Jeong,1 Bong-Hyuk Choi,1 Leif Hunter,2 Stuart Langley,3 Laszlo Pazmany,3 and Paul Trayhurn2 1School of Life and Food Sciences, Handong Global University, Pohang, Kyungbuk, Korea; 2Obesity Biology Unit, School of Clinical Sciences, University of Liverpool, University Clinical Departments, Liverpool L69 3GA, UK; 3University Hospital, Aintree, Clinical Sciences Centre, Longmoor Lane, Liverpool L7 9AL UK. We report here the use of human inflammation arrays to study the inflammatory gene expression profile of TNF-α- 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-α, and RNA from differentiated adipocytes with or without TNF- α treatment was hybridized to MWG human inflammation arrays to compare expression profiles. Eleven genes were up- or down-regulated in TNF-α-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-α. Taken together, we have identified several inflam- matory molecules expressed in SGBS adipocytes and dis- covered molecular factors explaining the relationship between obesity and atherosclerosis, focusing on inflammatory cyto- kines expressed in the TNF-α-treated SGBS cells. Further investigation into the role of these up- or down-regulated Received December 2, 2005 Accepted May 22, 2006 This work was supported by the Korea Research Foundation Grant (KRF-2003-013-C00089). Reprint address: requests to Dr. Myoung-Sool Do, School of Life and Food Sciences, Handong Global University Pohang, Korea. Tel: 82-54-260-1301, Fax: 82-54-261-1306, E-mail: msdo@ handong.edu cytokine genes during the pathological processes leading to the development of atherosclerosis is warranted. Key Words: Adipose tissue, cytokine, DNA inflammation, SGBS, TNF-α microarray, INTRODUCTION Adipose tissue is a storage organ for excess energy in the form of triglycerides. Since the dis- covery of leptin, numerous reports have been published, describing adipose tissue as a multi- functional organ that produces and secretes many factors, acting in a paracrine and endocrine manner.1 Adipocytes secrete many protein signals and factors, collectively known as adipocytokines or adipokines. These adipokines include inflam- matory molecules, such as leptin, resistin, adiponectin, tumor necrosis factor-α (TNF-α), adipsin, interleukins (IL-1β, IL-6, IL-8, IL-10), plasminogen activator inhibitor-1 (PAI-1), hapto- globin, C-reactive protein (CRP), and MCP-1.2,3 The current view is that certain adipokines and other factors produced and released by white adipose tissue are responsible for the chronic in- flammatory state of obesity, particularly the visceral form. Reduction of fat mass correlates with decreases in the serum levels of many of these adipokines,4 implying that approaches designed to promote fat loss should be useful in attenuating the proinflammatory milieu associated with obesity. The concomitance of inflammation, Yonsei Med J Vol. 47, No. 5, 2006