PARENT SESSION
Implantation & Early Development

MICROARRAY ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES INDUCED BY RELAXIN IN NORMAL HUMAN ENDOMETRIAL (NHE) STROMAL CELLS.

Deshpande, Usha¹, Lewis, Martyn¹, Unemori, Elaine¹, ¹

ABSTRACT- Relaxin, a 6 kDa pleiotropic hormone, is produced by the corpus luteum, the endometrium, and the placenta in humans. Relaxin has been shown to modulate the expression of several proteins in-vitro that may contribute to endometrial receptivity. There are several genes that are upregulated by Relaxin in endometrial stromal cells, and these include insulin-like growth factor binding protein-1, prolactin and HOXA-10. In addition, our previous work has shown that Relaxin stimulates the expression of two genes, vascular endothelial growth factor (VEGF) and cyclooxygenase (COX-2) in NHE cells. In an attempt to study the induction of other novel genes in response to Relaxin, we used cDNA microarrays to evaluate the gene expression pattern in NHE cells. Two fluorescent labeled cDNA probes were generated from the mRNA extracted from NHE cells that were either untreated or treated with Relaxin for 24 h. The two fluorescent probe samples were hybridized to a DNA microarray. Of the 9000 genes screened, the expression of several genes including structural proteins, transcription factors and cytokines were either upregulated or downregulated by Relaxin following 24 h stimulation. Of these, hepatocyte growth factor (HGF), an angiogenic factor, and tissue inhibitor of metalloproteinases (TIMP-3) were upregulated by 1.5 and 1.9 fold over the control treatment. The array results were validated by quantitative real time polymerase chain reaction (Q-PCR) and ELISA in separate studies. Thus, Relaxin may be important in regulating the vascular component of the endometrium via HGF, COX-2, and VEGF, and in maintaining endometrial integrity during the luteal phase via TIMP-3 regulation of extracellular matrix (ECM) remodeling.

KEY WORDS: Relaxin, Endometrium, Microarray, Angiogenic