Gene Expression Profiling in Human Preadipocytes and Adipocytes by Microarray Analysis

Urs, Sumithra; Smith, Colton; Campbell, Brett; Saxton, Arnold M.; Taylor, John; Zhang, Bing; Snoddy, Jay; Voy, Brynn Jones; Moustaid‐Moussa, Naima

doi:10.1093/jn/134.4.762

Cited by 119 publications

(81 citation statements)

References 48 publications

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“…40 It is likely that higher expression of SHMT2 in adipocytes is related to increased mitochondrial density. The relative small differences agree with previous studies 41,42 but may in part also be due to the selection of genes used, which was directed at white adipose characteristics and not to identify differences between white and brown adipose tissues. LiSa-2 and PAZ6 are both used to study adipocyte function, [8][9][10] but have a different physiological background.…”

Section: Discussionsupporting

confidence: 89%

Comparative expression analysis of isolated human adipocytes and the human adipose cell lines LiSa-2 and PAZ6

et al. 2008

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Objective: To obtain insight in the extent to which the human cell lines LiSa-2 and PAZ6 resemble isolated primary human adipocytes. Design: A combination of cDNA subtraction (representative difference analysis; RDA) and cDNA microarray analysis was used to select adipose specific genes to compare isolated (pre-)adipocytes with (un)differentiated LiSa-2 and PAZ6 cells. Measurements: RDA was performed on adipose tissue against lung tissue. A total of 1400 isolated genes were sequenced and cDNA microarray technology was used for further adipose related gene selection. 30 genes that were found to be enriched in adipose tissue were used to compare isolated human adipocytes and LiSa-2 and PAZ6 cells in the differentiated and undifferentiated states. Results: RDA and microarray analysis resulted in the identification of adipose enriched genes, but not in adipose specific genes. Of the 30 most differentially expressed genes, as expected, most were related to lipid metabolism. The second category consisted of methyltransferases, DNMT1, DNMT3a, RNMT and SHMT2, of which the expression was differentiation dependent and higher in differentiated adipocytes. Using the 30 adipose expressed genes, it was found that isolated adipocytes on one hand, and PAZ6 and LiSa-2 adipocytes on the other, differ primarily in lipid metabolism. Furthermore, LiSa-2 cells seem to be more similar to isolated adipocytes than PAZ6 cells. Conclusion:The LiSa-2 cell line is a good model for differentiated adipocytes, although one should keep in mind that the lipid metabolism in these cells deviates from the in vivo situation Furthermore, our results imply that methylation may have an important function in terminal adipocyte differentiation. In this study, we aimed to gain insight in the extent to which the human cell lines LiSa-2 and PAZ6 resemble human adipocytes in the in vivo situation, using adipose tissue properties. Therefore, we studied the expression levels of 30 adipose specific genes in isolated cells and in the undifferentiated and differentiated cell lines. Isolation of adipose tissue specific genes was attempted by using a combination of subtractive hybridization and microarray analysis.Subtractive hybridization, also named representation difference analysis (RDA), 16 is a method to enrich a target mRNA population, in this case mRNA from human subcutaneous adipose tissue, with low abundant and specific sequences, by removing those sequences that are also present in a second population, in this case lung mRNA. The resulting cDNA library was sequenced. The success of the subtraction procedure was assessed by spotting of the cDNA inserts on a microarray and by hybridizing this cDNA microarray with mRNA from different tissues, including adipose tissue and lung. By hybridization with RNA from adipose tissue and colon and lung tissue, 30 adipose enriched genes were selected. These were used for comparative analysis of isolated (pre-)adipocytes and (un)differentiated cell lines using cDNA microarray technology. During differentiation the medium...

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Section: Discussionsupporting

confidence: 89%

Comparative expression analysis of isolated human adipocytes and the human adipose cell lines LiSa-2 and PAZ6

et al. 2008

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“…41) It was also reported that expression of Smarcb1 was increased in human adipocytes compared with that in preadipocytes. 42) However, at the present, the functions of Ddx5 and Smarcb1 in adipogenesis, as well as that of glutathione S-transferase w1 (GSTw1), remain unclear. It was reported that HIF-3a is induced during differentiation of cardiac cells and alveolar type II cells.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia-Inducible Factor-3.ALPHA. Functions as an Accelerator of 3T3-L1 Adipose Differentiation

Hatanaka

Shimba

Sakaue

et al. 2009

Biological & Pharmaceutical Bulletin

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Adipose differentiation is a complex process associated with coordinated changes in gene expression, cell morphology, and hormone sensitivity.1) Several transcription factors influence these processes, among which peroxisome proliferator activated receptor g (PPARg) and CCAAT/enhancerbinding protein (C/EBP) family have been extensively studied.2,3) C/EBPb and C/EBPd are induced in the early stage of adipocyte differentiation and then mediate the expression of PPARg and C/EBPa. [4][5][6] In contrast to C/EBPd, C/EBPb is able to induce spontaneous differentiation in 3T3-L1 preadipocytes and enhance the adipogenic potential in NIH-3T3 fibroblasts. 5,6) Highly specific for adipose tissues, PPARg plays a critical role in the expression of most adipocyte-specific genes 7) and is able to convert nonadipogenic mesenchymal cells, such as fibroblasts and myoblasts, to adipocytes. 8,9) On the other hand, although developmentally necessary for adipogenesis, 10) C/EBPa is not always expressed during adipose differentiation and is critical in the establishment of insulin sensitivity. 11)Several recent studies have shown that hypoxia-inducible factor (HIF) is also involved in the regulation of adipose differentiation. [12][13][14] HIF is a transcriptional factor that plays a central role in oxygen homeostasis in mammalian cells by stimulating expression of oxygen-regulated genes related to erythropoiesis, angiogenesis, glucose uptake, and glycolysis. 15,16) HIF is a heterodimer composed of HIF-a and -b, both of which belong to basic helix-loop-helix (bHLH)/Per-ARNT-Sim (PAS) domain transcriptional factors.17) The HIFb subunit is identical to the arylhydrocarbon receptor nuclear translocator (ARNT) that also serves as a heterodimeric partner with the arylhydrocarbon receptor (AhR).18) In contrast, it appears that the HIF-a subunit's sole but critical function is to mediate the response to hypoxia. Under normoxic conditions, HIF-a subunits are hydroxylated at two conserved proline (Pro564 and Pro402 in HIF-1a) residues and one asparagine (Asn803 in HIF-1a) residue by HIF-a prolyl hydroxylases (PHD1, PHD2, and PHD3) and asaparagine hydroxylase FIH-1 (factor inhibiting HIF-1), respectively, and the hydroxylated HIF-a subunit proteins are destabilized and transcriptionally suppressed.19) Deprivation of oxygen by exposing cells to hypoxia prevents HIF hydroxylase activity and then stabilizes and transcriptionally activates HIF-a subunits. The first identified isoform of HIF-a, HIF-1a, was originally discovered as a high-affinity DNA-binding protein localized to the 3Ј hypoxia-responsive element (HRE) on the erythropoietin (EPO) gene. Using a subtraction cloning approach, Imagawa et al. found that HIF-1a mRNA is transiently induced in 3T3-L1 preadipocytes upon treatment with the adipogenic hormone cocktail containing insulin, dexamethasone, and 3-isobutyl-1-methylxanthine.12) Yun et al. found that HIF-1a represses PPARg2 gene expression and then inhibits adipogenesis through the induction of expression of transcriptional repressor DE...

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“…By using cDNA microarrays (25)(26)(27)(28)(29)(30)(31)(32) and proteomic technologies (Refs. 24 and 33 and references therein), it has been possible to identify several genes and proteins that are differentially regulated as a result of adipogenesis.…”

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confidence: 99%

“…In vivo transcript profiling studies of human and murine fat tissue (28,32,38) have shown the complexity of the adipocyte transcriptome and have implicitly established that the biology of these cells has a degree of intricacy that was not expected. To date, however, only two studies have investigated the proteome of human and mouse adipose tissue; one resolved about 100 human proteins using wide IPG strips and identified 16 polypeptides by means of mass spectrometry (39), while the other resolved a considerable number of murine white adipose tissue proteins and identified 80 unique polypeptides (40).…”

mentioning

confidence: 99%

Identification of Extracellular and Intracellular Signaling Components of the Mammary Adipose Tissue and Its Interstitial Fluid in High Risk Breast Cancer Patients

Celis

Moreira

Cabezón

et al. 2005

Molecular & Cellular Proteomics

208

164

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It has become clear that growth and progression of breast tumor cells not only depend on their malignant potential but also on factors present in the tumor microenvironment. Of the cell types that constitute the mammary stroma, the adipocytes are perhaps the least well studied despite the fact that they represent one of the most prominent cell types surrounding the breast tumor cells. There is compelling evidence demonstrating a role for the mammary fat pad in mammary gland development, and some studies have revealed the ability of fat tissue to augment the growth and ability to metastasize of mammary carcinoma cells. Very little is known, however, about which factors adipocytes produce that may orchestrate these actions and how this may come about. In an effort to shed some light on these questions, we present here a detailed proteomic analysis, using two-dimensional gel-based technology, mass spectrometry, immunoblotting, and antibody arrays, of adipose cells and interstitial fluid of fresh fat tissue samples collected from sites topologically distant from the tumors of high risk breast cancer patients that underwent mastectomy and that were not treated prior to surgery. A total of 359 unique proteins were identified, including numerous signaling molecules, hormones, cytokines, and growth factors, involved in a variety of biological processes such as signal transduction and cell communication; energy metabolism; protein metabolism; cell growth and/or maintenance; immune response; transport; regulation of nucleobase, nucleoside, and nucleic acid metabolism; and apoptosis. Apart from providing a comprehensive overview of the mammary fat proteome and its interstitial fluid, the results offer some insight as to the role of adipocytes in the breast tumor microenvironment and provide a first glance of their molecular cellular circuitry. In addition, the results open new possibilities to the study of obesity, which has a strong association with type 2 diabetes, hypertension, and coronary heart disease. Molecular & Cellular Proteomics 4: 492-522, 2005.During the last years there have been numerous reports indicating that growth and progression of breast as well as other tumor cells depend not only on their malignant potential but also on stromal factors present in the tumor microenvironment, the insoluble extracellular matrix as well as cell-cell interactions (Refs.

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Gene Expression Profiling in Human Preadipocytes and Adipocytes by Microarray Analysis

Cited by 119 publications

References 48 publications

Comparative expression analysis of isolated human adipocytes and the human adipose cell lines LiSa-2 and PAZ6

Comparative expression analysis of isolated human adipocytes and the human adipose cell lines LiSa-2 and PAZ6

Hypoxia-Inducible Factor-3.ALPHA. Functions as an Accelerator of 3T3-L1 Adipose Differentiation

Identification of Extracellular and Intracellular Signaling Components of the Mammary Adipose Tissue and Its Interstitial Fluid in High Risk Breast Cancer Patients

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