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Progesterone up-regulates WT1 mRNA and protein, and alters the relative expression of WT1 transcripts in cultured endometrial stromal cells

Progesterone up-regulates WT1 mRNA and protein, and alters the relative expression of WT1 transcripts in cultured endometrial stromal cells
Progesterone up-regulates WT1 mRNA and protein, and alters the relative expression of WT1 transcripts in cultured endometrial stromal cells
Objective
To determine the change in expression of the Wilms tumor suppressor gene product, WT1, by progesterone alone in endometrial stromal cell culture and to study its relationship with prolactin, a marker of decidualization. In addition, to examine the change in ratio of WT1 isoforms with and without exon 5 message.
Methods
Endometrial biopsies were taken from eight patients who had hysterectomy. Stromal cells were isolated and cultured in the presence of progesterone alone (12 days) or progesterone and 8-bromo-cyclic adenosine monophosphate (cAMP) (6 days). RNA was extracted from cells, and reverse transcription, real-time polymerase chain reaction (PCR), and conventional PCR were done to analyze WT1 mRNA expression. Immunocytochemistry was performed on equivalent cells to study WT1 protein expression. Decidualization was identified by increased prolactin concentrations in the media and immunocytochemical markers IGFBP-1 and collagen IV.
Results
Reverse transcription and real-time PCR revealed a significant increase in WT1 mRNA with increasing progesterone concentrations when decidualization was occurring (n = 6, P = .002). Increasing progesterone concentrations also increased the proportion of the WT1 transcript containing a 17-amino-acid insert (+ exon 5 expression); changes in WT1 exon 5 expression have been shown to be involved in control of proliferation and differentiation. Significant correlations between WT1 message and prolactin existed at physiologic progesterone concentrations (6.25, 12.5, 25, and 50 nM; P < .05) until prolactin concentrations reached a plateau at 100 nM. At concentrations of progesterone alone (> 25 nM) and progesterone with 8-bromo-cAMP, WT1 protein was localized to the nuclei of many of the decidualized stromal cells.
Conclusion
The changing expression of WT1 isoforms in endometrial stromal cells caused by progesterone may be important for differentiation into the decidualized phenotype.
decidualization, endometrium, progesterone, wt1
1071-5576
509-516
Anthony, F.W.
398b8bed-237e-4fad-a77d-89266fb00bc0
Mukhtar, D.D.
4a16db08-521a-4359-9991-b345570f281c
Pickett, M.A.
4b358cb6-2f2e-4691-a314-0e80df48eadb
Cameron, I.T.
f7595539-efa6-4687-b161-e1e93ff710f2
Anthony, F.W.
398b8bed-237e-4fad-a77d-89266fb00bc0
Mukhtar, D.D.
4a16db08-521a-4359-9991-b345570f281c
Pickett, M.A.
4b358cb6-2f2e-4691-a314-0e80df48eadb
Cameron, I.T.
f7595539-efa6-4687-b161-e1e93ff710f2

Anthony, F.W., Mukhtar, D.D., Pickett, M.A. and Cameron, I.T. (2003) Progesterone up-regulates WT1 mRNA and protein, and alters the relative expression of WT1 transcripts in cultured endometrial stromal cells. Journal of the Society for Gynecologic Investigation, 10 (8), 509-516. (doi:10.1016/S1071-5576(03)00147-3).

Record type: Article

Abstract

Objective
To determine the change in expression of the Wilms tumor suppressor gene product, WT1, by progesterone alone in endometrial stromal cell culture and to study its relationship with prolactin, a marker of decidualization. In addition, to examine the change in ratio of WT1 isoforms with and without exon 5 message.
Methods
Endometrial biopsies were taken from eight patients who had hysterectomy. Stromal cells were isolated and cultured in the presence of progesterone alone (12 days) or progesterone and 8-bromo-cyclic adenosine monophosphate (cAMP) (6 days). RNA was extracted from cells, and reverse transcription, real-time polymerase chain reaction (PCR), and conventional PCR were done to analyze WT1 mRNA expression. Immunocytochemistry was performed on equivalent cells to study WT1 protein expression. Decidualization was identified by increased prolactin concentrations in the media and immunocytochemical markers IGFBP-1 and collagen IV.
Results
Reverse transcription and real-time PCR revealed a significant increase in WT1 mRNA with increasing progesterone concentrations when decidualization was occurring (n = 6, P = .002). Increasing progesterone concentrations also increased the proportion of the WT1 transcript containing a 17-amino-acid insert (+ exon 5 expression); changes in WT1 exon 5 expression have been shown to be involved in control of proliferation and differentiation. Significant correlations between WT1 message and prolactin existed at physiologic progesterone concentrations (6.25, 12.5, 25, and 50 nM; P < .05) until prolactin concentrations reached a plateau at 100 nM. At concentrations of progesterone alone (> 25 nM) and progesterone with 8-bromo-cAMP, WT1 protein was localized to the nuclei of many of the decidualized stromal cells.
Conclusion
The changing expression of WT1 isoforms in endometrial stromal cells caused by progesterone may be important for differentiation into the decidualized phenotype.

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Published date: December 2003
Keywords: decidualization, endometrium, progesterone, wt1

Identifiers

Local EPrints ID: 25200
URI: http://eprints.soton.ac.uk/id/eprint/25200
ISSN: 1071-5576
PURE UUID: 00d16d12-b047-479c-856b-6c7265664765
ORCID for I.T. Cameron: ORCID iD orcid.org/0000-0002-4875-267X

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Date deposited: 07 Apr 2006
Last modified: 16 Mar 2024 03:00

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Contributors

Author: F.W. Anthony
Author: D.D. Mukhtar
Author: M.A. Pickett
Author: I.T. Cameron ORCID iD

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