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


CONCLUSIONS
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.
Leuven University Fertility Centre, Department of Obstetrics & Gynaecology, University Hospital Gasthuisberg, Leuven, Belgium.
Amelie
A
Fassbender
Peter
P
Simsa
Cleophas M
CM
Kyama
Etienne
E
Waelkens
Attila
A
Mihalyi
Christel
C
Meuleman
Olivier
O
Gevaert
Raf
R
Van de Plas
Bart
B
de Moor
Thomas M
TM
D'Hooghe
D000328:Adult; D016022:Case-Control Studies; D002458:Cell Fractionation / Q000379:methods; D004715:Endometriosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004717:Endometrium / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D006146:Guanidines / Q000494:pharmacology*; D006801:Humans; D008183:Luteal Phase / Q000235:genetics / Q000378:metabolism; D046228:Microarray Analysis / Q000379:methods*; D010636:Phenols / Q000494:pharmacology*; D020543:Proteome / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D013048:Specimen Handling / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014591:Uterine Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology
D006146:Guanidines; D010636:Phenols; D020543:Proteome; D012333:RNA, Messenger; C411644:trizol