Progesterone receptor antagonists Org 31710 and RU 486 increase apoptosis in human periovulatory granulosa cells.
OBJECTIVE
To investigate if progesterone receptor (PR)-mediated effects are involved in regulating the susceptibility to apoptosis in LH receptor-stimulated human luteinizing granulosa cells.


METHODS
Laboratory study.


METHODS
Göteborg University and an in vitro fertilization laboratory of a university hospital.


METHODS
Women undergoing oocyte retrieval for in vitro fertilization after ovulation induction with gonadotropins.


METHODS
Luteinizing granulosa cells were isolated from follicular aspirates after oocyte removal. The cells were treated with or without RU 486 (1 microM-100 microM), Org 31710 (1 microM-100 microM), progesterone (1 nM-10 microM), dexamethasone (0.5 microM-100 microM), dihydrotestosterone (1 nM-25 microM), RU 486 (10 microM-100 microM) + dexamethasone (50 microM), and picrotoxin (1 microM-100 microM) and were cultured under serum-free conditions.


METHODS
Measurement of caspase-3 activity; detection of internucleosomal DNA fragmentation using gel electrophoresis and fluorospectrophotometry; progesterone analysis of spent medium.


RESULTS
Addition of the PR antagonists RU 486 or Org 31710 in vitro to human luteinizing granulosa cells caused an increase in caspase-3 activity and a dose-dependent increase in internucleosomal DNA fragmentation. No effect on DNA fragmentation was seen after addition of dexamethasone, dihydrotestosterone, or picrotoxin.


CONCLUSIONS
Nuclear PR-mediated effects are involved in regulating the susceptibility to apoptosis in LH receptor-stimulated human luteinizing granulosa cells.
Svensson
E C
EC
Department of Physiology, Göteborg University, Göteborg, Sweden.
Markström
E
E
Shao
R
R
Andersson
M
M
Billig
H
H
D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects*; D003907:Dexamethasone / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D004963:Estrenes / Q000494:pharmacology*; D005260:Female; D005663:Furans / Q000494:pharmacology*; D018756:GABA Antagonists / Q000494:pharmacology; D005938:Glucocorticoids / Q000494:pharmacology; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects*; D006727:Hormone Antagonists / Q000494:pharmacology*; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology*; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism; D010852:Picrotoxin / Q000494:pharmacology; D011374:Progesterone / Q000493:pharmacokinetics / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000172:deficiency; D011980:Receptors, Progesterone / Q000037:antagonists & inhibitors*; D013050:Spectrometry, Fluorescence
D053148:Caspase 3; D020169:Caspases; D003907:Dexamethasone; D013196:Dihydrotestosterone; D004963:Estrenes; D005663:Furans; D018756:GABA Antagonists; D005938:Glucocorticoids; D006727:Hormone Antagonists; D015735:Mifepristone; D009707:Nucleosomes; D010852:Picrotoxin; D011374:Progesterone; D011963:Receptors, GABA-A; D011980:Receptors, Progesterone; C076196:Org 31710; C505407:CASP3 protein, human