Progestin stimulates the biosynthesis of fibronectin and accumulation of fibronectin mRNA in human endometrial stromal cells.
Fibronectin is a major component of decidual basement membrane. In the present study, we have investigated the effect of progestin on the synthesis and secretion of fibronectin in human endometrial stromal cells. Stromal cells were isolated during the menstrual cycle and cultured in RPMI-1640 with 2% fetal calf serum supplemented with progesterone or medroxyprogesterone acetate (MPA) in a long-term culture system. Indirect immunofluorescent staining showed that fibronectin was uniformly distributed in the intracellular and extracellular regions of stromal cells treated with MPA for 14 days. The biosynthesis and secretion of this protein and the accumulation of cellular fibronectin mRNA were studied after various culture periods. Cells were pulse-labelled with [35S]methionine to determine the amount of newly synthesized fibronectin secreted into the culture medium. A monoclonal antibody (Mab) identified human fibronectin on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), showing a predominant band (Mr 230-250 kDa) which migrated with authentic fibronectin run in parallel. In six endometrial specimens, the amount of radioactivity incorporated as [35S]fibronectin was increased by progestin. Maximal stimulation occurred after 6 days treatment with MPA. Culture beyond 16 days reduced the rate of synthesis and secretion to 40% of the maximum. The effect of progestin was dose dependent with 0.02, 0.2 and 1 microM progesterone, producing 2.0, 3.8 and 11-fold increases respectively, over the control. Medroxyprogesterone acetate was more effective than progesterone, the maximal response (10-fold increase) being achieved at 0.02 microM MPA.(ABSTRACT TRUNCATED AT 250 WORDS)
Zhu
H H
HH
Department of Obstetrics and Gynecology, School of Medicine, State University of New York, Stony Brook 11794.
Huang
J R
JR
Mazela
J
J
Elias
J
J
Tseng
L
L
D002478:Cells, Cultured / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004717:Endometrium / Q000187:drug effects*; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D008525:Medroxyprogesterone / Q000031:analogs & derivatives* / Q000494:pharmacology; D017258:Medroxyprogesterone Acetate; D015735:Mifepristone / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D012065:Relaxin / Q000494:pharmacology
D005353:Fibronectins; D012333:RNA, Messenger; D015735:Mifepristone; D012065:Relaxin; D017258:Medroxyprogesterone Acetate; D008525:Medroxyprogesterone