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

[1] INSULIN STIMULATES GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE GENE-EXPRESSION THROUGH CIS-ACTING DNA-SEQUENCES [J].

ALEXANDER, MC ;

LOMANTO, M ;

NASRIN, N ;

RAMAIKA, C .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (14) :5092-5096

[2] GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues [J].

Barber, RD ;

Harmer, DW ;

Coleman, RA ;

Clark, BJ .

PHYSIOLOGICAL GENOMICS, 2005, 21 (03) :389-395

[3] Norepinephrine, tri-iodothyronine and insulin upregulate glyceraldehyde-3-phosphate dehydrogenase mRNA during brown adipocyte differentiation [J].

Barroso, I ;

Benito, B ;

García-Jiménez, C ;

Hernández, A ;

Obregón, MJ ;

Santisteban, P .

EUROPEAN JOURNAL OF ENDOCRINOLOGY, 1999, 141 (02) :169-179

[4] Local androgen inactivation in abdominal visceral adipose tissue [J].

Blouin, K ;

Richard, C ;

Bélanger, C ;

Dupont, P ;

Daris, M ;

Laberge, P ;

Luu-The, V ;

Tchernof, A .

JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 2003, 88 (12) :5944-5950

[5] Potential involvement of adipocyte insulin resistance in obesity-associated up-regulation of adipocyte lysophospholipase D/autotaxin expression [J].

Boucher, J ;

Quilliot, D ;

Pradères, JP ;

Simon, MF ;

Grés, S ;

Guigné, C ;

Prévot, D ;

Ferry, G ;

Boutin, JA ;

Carpéné, C ;

Valet, P ;

Saulnier-Blache, JS .

DIABETOLOGIA, 2005, 48 (03) :569-577

[6] Quantitative real-time RT-PCR - a perspective [J].

Bustin, SA ;

Benes, V ;

Nolan, T ;

Pfaffl, MW .

JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2005, 34 (03) :597-601

[7] Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [J].

Bustin, SA .

JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2000, 25 (02) :169-193

[8] Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems [J].

Bustin, SA .

JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2002, 29 (01) :23-39

[9] Variation in the calpain-10 gene is associated with elevated triglyceride levels and reduced adipose tissue messenger ribonucleic acid expression in obese Swedish subjects [J].

Carlsson, E ;

Fredriksson, J ;

Groop, L ;

Ridderstråle, M .

JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 2004, 89 (07) :3601-3605

[10] ADIPOSE-TISSUE-SPECIFIC INCREASE IN GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE-ACTIVITY AND MESSENGER-RNA AMOUNTS IN SUCKLING PRE-OBESE ZUCKER RATS - EFFECT OF WEANING [J].

DUGAIL, I ;

QUIGNARDBOULANGE, A ;

BAZIN, R ;

LELIEPVRE, X ;

LAVAU, M .

BIOCHEMICAL JOURNAL, 1988, 254 (02) :483-487

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