Validation of endogenous control genes in human adipose tissue:: Relevance to obesity and obesity-associated type 2 diabetes mellitus

被引：95

作者：

Catalan, V. ^{[1
]}

Gomez-Ambrosi, J. ^{[1
]}

Rotellar, F. ^{[2
]}

Silva, C. ^{[3
]}

Rodriguez, A. ^{[1
]}

Salvador, J. ^{[3
]}

Gil, M. J. ^{[4
]}

Cienfuegos, J. A. ^{[2
]}

Fruhbeck, G. ^{[1
,3
]}

机构：

[1] Univ Navarra, Clin Univ Navarra, Metab Res Lab, Pamplona 31008, Spain

[2] Univ Navarra, Clin Univ Navarra, Dept Surg, Pamplona 31008, Spain

[3] Univ Navarra, Clin Univ Navarra, Dept Endocrinol, Pamplona 31008, Spain

[4] Univ Navarra, Clin Univ Navarra, Dept Biochem, Pamplona 31008, Spain

来源：

HORMONE AND METABOLIC RESEARCH | 2007年 / 39卷 / 07期

关键词：

obesity; type 2 diabetes mellitus; real-time PCR; adipose tissue; housekeeping gene expression; POLYMERASE CHAIN-REACTION; MESSENGER-RNA; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; RIBOSOMAL-RNA; BETA-ACTIN; HOUSEKEEPING GENES; INTERNAL STANDARDS; RT-PCR; EXPRESSION; ADIPOCYTE;

D O I：

10.1055/s-2007-982502

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

The aim of the present study was to test the influence of obesity and the presence of type 2 diabetes mellitus (T2DM) on the expression of ten housekeeping genes and of the 18S rRNA in a group of human adipose tissue samples from the omental and subcutaneous depot. Adipose tissue biopsies were obtained by laparoscopic surgery from lean and obese patients. After the extraction, mRNA levels in adipose tissue samples were quantified by real-time PCR using the commercial Human Endogenous Control Plates. From the genes analyzed, 18S rRNA exhibited the most stable expression levels in both depots regardless of the pathophysiological conditions of obesity and obesity-associated T2DM. Contrarily, GAPD was the gene with the highest variation in its expression levels, being upregulated (8.0-fold) in the obese group and downregulated (3.5-fold) in obesity-associated T2DM. Our results show that 18S rRNA may be the most suitable gene for normalization in expression studies performed in human adipose tissue samples obtained from patients suffering from obesity and/or obesity-associated T2DM, whereas GAPD is less appropriate for comparison purposes under these circumstances.

引用

页码：495 / 500

页数：6

共 58 条

[51] Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes [J].

Vandesompele, Jo ;

De Preter, Katleen ;

Pattyn, Filip ;

Poppe, Bruce ;

Van Roy, Nadine ;

De Paepe, Anne ;

Speleman, Frank .

GENOME BIOLOGY, 2002, 3 (07)

[52] Adipose tissue gene expression in obese subjects during low-fat and high-fat hypocaloric diets [J].

Viguerie, N ;

Vidal, H ;

Arner, P ;

Holst, C ;

Verdich, C ;

Avizou, S ;

Astrup, A ;

Saris, WHM ;

Macdonald, IA ;

Klimcakova, E ;

Clément, K ;

Martinez, A ;

Hoffstedt, J ;

Sorensen, TIA ;

Langin, D .

DIABETOLOGIA, 2005, 48 (01) :123-131

[53] A survey of genes differentially expressed in subcutaneous and visceral adipose tissue in men [J].

Vohl, MC ;

Sladek, R ;

Robitaille, J ;

Gurd, S ;

Marceau, P ;

Richard, D ;

Hudson, TJ ;

Tchernof, A .

OBESITY RESEARCH, 2004, 12 (08) :1217-1222

[54] Comparison of human adult and fetal expression and identification of 535 housekeeping/maintenance genes [J].

Warrington, JA ;

Nair, A ;

Mahadevappa, M ;

Tsyganskaya, M .

PHYSIOLOGICAL GENOMICS, 2000, 2 (03) :143-147

[55] Precision of measurement and body size in whole-body air-displacement plethysmography [J].

Wells, JCK ;

Fuller, NJ .

INTERNATIONAL JOURNAL OF OBESITY, 2001, 25 (08) :1161-1167

[56] Adiponectin - a key adipokine in the metabolic syndrome [J].

Whitehead, JP ;

Richards, AA ;

Hickman, IJ ;

Macdonald, GA ;

Prins, JB .

DIABETES OBESITY & METABOLISM, 2006, 8 (03) :264-280

[57] Direct comparison of GAPDH, β-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia [J].

Zhong, H ;

Simons, JW .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 259 (03) :523-526

[58] Fudenine, a C-terminal truncated rat homologue of mouse prominin, is blood glucose-regulated and can up-regulate the expression of GAPDH [J].

Zhu, GZ ;

Chang, YS ;

Zuo, J ;

Dong, XY ;

Zhang, M ;

Hu, GX ;

Fang, FD .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2001, 281 (04) :951-956

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