Mechanisms of matrix metalloproteinase-2 (mmp-2) transcriptional repression by progesterone in jar choriocarcinoma cells.
BACKGROUND
Although the MMP-2 promoter lacks a canonical progesterone response element (PRE), the hormone inhibits MMP-2 expression and is part of treatment protocols in gynecological invasive pathologies, including endometriosis and endometrial hyperplasia. This study aimed to explore the mechanism by which progesterone inhibits MMP-2 expression.


METHODS
The effect of progesterone on MMP-2 expression in the JAR human choriocarcinoma cell line was analyzed by gelatin zymography. MMP-2 transcript expression was studied using Northern blot and semi-quantitative RT-PCR. Rat promoter deletion analysis, electrophoretic mobility shift and chromatin immuno-precipitation assays were performed in order to locate the DNA binding site and the transcription factors involved in MMP-2 regulation.


RESULTS
Progesterone significantly decreased secretion of pro-MMP-2 and MMP-2 transcript expression level in a dose-dependent manner. Progesterone (1 microM) significantly decreased both human and rat MMP-2 promoter activity (80.1% +/- 0.3 and 81.3% +/- 0.23, respectively). Progesterone acts through the SP1 family transcription factors-binding site, located between -1433 and -1342 bp region from the transcriptional start site of the rat MMP-2 promoter, which are present in the orthologous human MMP-2 promoter. Progesterone receptor (PR), SP2, SP3 and SP4 proteins are constitutively bound to this consensus sequence.


CONCLUSIONS
Progesterone reduces PR and SP4 binding to the MMP-2 promoter, thereby suppressing transcription. Progesterone also promotes SP4 degradation. These novel mechanisms of MMP-2 regulation by progesterone provide the biological rationale for the use of progesterone in clinical settings associated with increased MMP-2 expression.
Goldman
Shlomit
S
Laboratory for Research in Reproductive Sciences, Department of Obstetrics and Gynecology, HaEmek Medical Centre, Afula, Israel. goldman_s@clalit.org.il
Lovett
David H
DH
Shalev
Eliezer
E
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002822:Choriocarcinoma / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011374:Progesterone / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D051381:Rats; D011980:Receptors, Progesterone / Q000378:metabolism; D020218:Response Elements / Q000502:physiology; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051706:Sp4 Transcription Factor / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology; D014594:Uterine Neoplasms / Q000235:genetics* / Q000378:metabolism*
D011980:Receptors, Progesterone; C497124:SP4 protein, human; D016329:Sp1 Transcription Factor; D051706:Sp4 Transcription Factor; D011374:Progesterone; D020778:Matrix Metalloproteinase 2