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This version published online on May 11, 2006
Endocrinology, doi:10.1210/en.2006-0167
A more recent version of this article appeared on August 1, 2006
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Submitted on February 9, 2006
Accepted on May 4, 2006

FOXO1A DIFFERENTIALLY REGULATES GENES OF DECIDUALIZATION

Oscar L. Buzzio, Zhenxiao Lu, Curt D. Miller, Terry G. Unterman, and J. Julie Kim*

Department of Obstetrics and Gynecology, Northwestern University, Chicago, IL, 60611, Departments of Physiology and Biophysics, and Medicine, University of Illinois at Chicago, College of Medicine and VA Chicago Healthcare System (West Side), Chicago, IL, 60612

* To whom correspondence should be addressed. E-mail: j-kim4{at}northwestern.edu.

FOXO1A has been identified as one gene that is upregulated early in the decidualization process. To further investigate the role of FOXO1A during this process, 6 genes IGFBP1, PRL, TIMP3, LAMB1, CNR1, and DCN, shown to be upregulated during decidualization, were chosen as potential targets of FOXO1A action. Treatment of human endometrial stromal cells (HSC) with the hormones (estradiol, medroxyprogesterone acetate) plus dibutyryl cAMP (H+dbcAMP) for 48 h increased expression of IGFBP1, PRL, TIMP3, CNR1 and DCN, but not LAMB1, as measured by real-time PCR. Silencing of FOXO1A using siRNA oligonucleotides, decreased IGFBP1, and DCN levels, and increased CNR1, TIMP3 and PRL levels. LAMB1 was not affected. When FOXO1A was overexpressed in HSC, expression of IGFBP1, DCN and PRL increased, while levels of TIMP3 and CNR1 decreased. Addition of H+dbcAMP caused an increased expression of IGFBP1, PRL and DCN beyond that of FOXO1A alone. TIMP3 and CNR1 levels decreased even further in response to H+dbcAMP compared with FOXO1A alone. LAMB1, which was unresponsive to FOXO1A, decreased when H+dbcAMP was added. Overexpressing FOXO1A also caused a change in cell shape, in that the stromal fibroblasts acquired a rounded, epithelioid appearance. Finally, reporter studies showed that co-transfection of FOXO1A significantly increased PRL promoter activity but not TIMP3 promoter activity. Addition of H+dbcAMP resulted in a significant increase in PRL promoter activity and a significant decrease in TIMP3 promoter activity. In summary, this study demonstrates the versatile nature of FOXO1A in the regulation of a number of decidualization-specific genes.
Key words: decidualization • FOXO1A • endometrium




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