Open Access Methodology

TRIzol treatment of secretory phase endometrium allows combined proteomic and mRNA microarray analysis of the same sample in women with and without endometriosis

Amelie Fassbender1, Peter Simsa1, Cleophas M Kyama12, Etienne Waelkens3, Attila Mihalyi1, Christel Meuleman1, Olivier Gevaert4, Raf Van de Plas4, Bart de Moor4 and Thomas M D'Hooghe12*

Author Affiliations

1 Leuven University Fertility Centre, Department of Obstetrics & Gynaecology, University Hospital Gasthuisberg, Leuven, Belgium

2 Division of Reproductive Health and Biology, Institute of Primate Research, P.O. Box 24481-00502 Karen, Nairobi, Kenya

3 Biochemistry Section, Department of Molecular Cell Biology, Campus Gasthuisberg, Leuven, Belgium

4 Department of Electrical Engineering, ESAT-SCD, K.U.Leuven, Kasteelpark-Arenberg 10, B-3001 Heverlee, Belgium

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Reproductive Biology and Endocrinology 2010, 8:123  doi:10.1186/1477-7827-8-123

Published: 21 October 2010

Abstract

Background

According to mRNA microarray, proteomics and other studies, biological abnormalities of eutopic endometrium (EM) are involved in the pathogenesis of endometriosis, but the relationship between mRNA and protein expression in EM is not clear. We tested for the first time the hypothesis that EM TRIzol extraction allows proteomic Surface Enhanced Laser Desorption/Ionisation Time-of-Flight Mass Spectrometry (SELDI-TOF MS) analysis and that these proteomic data can be related to mRNA (microarray) data obtained from the same EM sample from women with and without endometriosis.

Methods

Proteomic analysis was performed using SELDI-TOF-MS of TRIzol-extracted EM obtained during secretory phase from patients without endometriosis (n = 6), patients with minimal-mild (n = 5) and with moderate-severe endometriosis (n = 5), classified according to the system of the American Society of Reproductive Medicine. Proteomic data were compared to mRNA microarray data obtained from the same EM samples.

Results

In our SELDI-TOF MS study 32 peaks were differentially expressed in endometrium of all women with endometriosis (stages I-IV) compared with all controls during the secretory phase. Comparison of proteomic results with those from microarray revealed no corresponding genes/proteins.

Conclusion

TRIzol treatment of secretory phase EM allows combined proteomic and mRNA microarray analysis of the same sample, but comparison between proteomic and microarray data was not evident, probably due to post-translational modifications.