Progestin represses human connexin43 gene expression similarly in primary cultures of myometrial and uterine leiomyoma cells.

Abstract

The mechanism by which progestin represses the expression of the human connexin43 (cx43) gene was analyzed in primary cultures of human myometrial cells, and for comparison, in primary cultures of uterine leiomyoma cells. Within 24 h, the levels of connexin43 (Cx43) protein in primary cells treated with progestin were reduced to about 50% of that in untreated cells, and these levels were maintained for up to 120 h. A plateau in the reduction of Cx43 protein levels was reached at progestin concentrations of 50-100 nM. No significant difference was found in a comparison of progestin-mediated reduction of Cx43 protein in autologous myometrial and leiomyoma primary cultures. Levels of cx43 mRNA levels also decreased to about 50% in myometrial and leiomyoma cells within hours after treatment with progestin, and these new levels were maintained for up to 48 h. Nuclear run-on transcription analysis showed that 100 nM progestin partially repressed transcription of the cx43 gene in both myometrial and leiomyoma primary cultures. The amount of decrease in cx43 transcription in cells treated with progestin was paralleled by a corresponding decrease in cytoplasmic cx43 mRNA levels. Progesterone receptor (PR)-mediated transcription was also determined to be similar in the two types of primary cells as evidenced by transient expression assays. Thus progestin down-regulates the expression of the human cx43 gene primarily by repressing transcription of the gene in myometrial cells, and it acts similarly in leiomyoma cells.