Comparison in gene expression of secretory human endometrium using laser microdissection.
BACKGROUND
The endometrium prepares for implantation under the control of steroid hormones. It has been suggested that there are complicated interactions between the epithelium and stroma in the endometrium during menstrual cycle. In this study, we demonstrate a difference in gene expression between the epithelial and stromal areas of the secretory human endometrium using microdissection and macroarray technique.


METHODS
The epithelial and stromal areas were microdissected from the human endometrium during the secretory phase. RNA was extracted and amplified by PCR. Macroarray analysis of nearly 1000 human genes was carried out in this study. Some genes identified by macroarray analysis were verified using real-time PCR.


RESULTS
In this study, changes in expression <2.5-fold in three samples were excluded. A total of 28 genes displayed changes in expression from array data. Fifteen genes were strongly expressed in the epithelial areas, while 13 genes were strongly expressed in the stromal areas. The strongly expressed genes in the epithelial areas with a changes >5-fold were WAP four-disulfide core domain 2 (44.1 fold), matrix metalloproteinase 7 (40.1 fold), homeo box B5 (19.8 fold), msh homeo box homolog (18.8 fold), homeo box B7 (12.7 fold) and protein kinase C, theta (6.4 fold). On the other hand, decorin (55.6 fold), discoidin domain receptor member 2 (17.3 fold), tissue inhibitor of metalloproteinase 1 (9 fold), ribosomal protein S3A (6.3 fold), and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains (5.2 fold) were strongly expressed in the stromal areas. WAP four-disulfide core domain 2 (19.4 fold), matrix metalloproteinase 7 (9.7-fold), decorin (16.3-fold) and tissue inhibitor of metalloproteinase 1 (7.2-fold) were verified by real-time PCR.


CONCLUSIONS
Some of the genes we identified with differential expression are related to the immune system. These results are telling us the new information for understanding the secretory human endometrium.
Yanaihara
Atsushi
A
Department of Obstetrics and Gynecology, Showa University School of Medicine, Hatanodai, Shinagawa-ku, Tokyo, Japan. atsushi-y@med.showa-u.ac.jp
Otsuka
Yukiko
Y
Iwasaki
Shinji
S
Koide
Keiko
K
Aida
Tadateru
T
Okai
Takashi
T
D003199:Computer Systems; D000070818:Discoidin Domain Receptors; D004717:Endometrium / Q000737:chemistry* / Q000378:metabolism*; D030501:Epididymal Secretory Proteins / Q000235:genetics; D004847:Epithelial Cells / Q000737:chemistry / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006801:Humans; D007834:Lasers; D008183:Luteal Phase / Q000235:genetics*; D020783:Matrix Metalloproteinase 7; D008666:Metalloendopeptidases / Q000235:genetics; D042282:Microdissection / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D011975:Receptors, Mitogen / Q000235:genetics; D017154:Stromal Cells / Q000737:chemistry / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D023083:beta-Defensins
C426184:DEFB126 protein, human; D030501:Epididymal Secretory Proteins; D011975:Receptors, Mitogen; D019715:Tissue Inhibitor of Metalloproteinase-1; D023083:beta-Defensins; D012313:RNA; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases; D008666:Metalloendopeptidases; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7