Endometrial effects of RU486 in primates--antiproliferative action despite signs of estrogen action and increased cyclin-B expression.
Continuous antiprogestin administration to hormone replaced, castrate monkeys inhibits estrogen-induced endometrial proliferation through mechanisms which remains unclear. To elucidate the molecular mechanisms of RU486-induced endometrial suppression, we treated six intact female cynomolgus monkeys on cycle days 2-22 sequentially with placebo, RU486 (1 mg/kg/day) and levonorgestrel (LNG) (2 microg/kg/day) intramuscularly (i.m.), with uterine wedge sections and endometrial biopsies collected on day 22 of each cycle. The uterine sections were evaluated for morphology, mitosis and proliferating cell nuclear antigen (PCNA) immunohistochemistry. Changes in the mRNA levels of ER, PR, cyclin-B and tumour suppressor gene p21 were assessed using co-amplification with beta-actin by reverse transcriptase-polymerase chain reaction (RT-PCR). Administration of RU486 uniformly resulted in characteristic suppression of endometrium with few mitosis, dense stroma and simple glands, whereas the effects of LNG were less uniform. Following RU486 administration, the levels of endometrial ER and PR mRNA were comparable to proliferative phase endometrium, and significantly higher than those seen in the secretory endometrium, indicating that some of the biological actions of E2 were not inhibited during RU486 treatment. Despite scarce mitosis, PCNA was readily detectable in all samples. Curiously, in comparison to secretory phase controls, the levels of cyclin-B, but not p21, mRNA were markedly increased following RU486. The effects of LNG on the levels of these mRNA species varied, with mean levels falling between those of the secretory phase controls, and RU486-treated specimens. The increase in cyclin-B mRNA and lack of mitosis suggests that anti-proliferative actions of RU486 in the primate endometrium might be associated with a cell-cycle block at the G2-M interphase. Whether mechanisms similar to these are associated with the beneficial clinical effects of RU486 seen in the treatment of various hormone dependent maladies remains to be determined.
Heikinheimo
O
O
The Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, VA 23517, USA.
Hsiu
J G
JG
Gordon
K
K
Kim
S
S
Williams
R F
RF
Gibbons
W E
WE
Hodgen
G D
GD
D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D016213:Cyclins / Q000096:biosynthesis*; D017931:DNA Primers; D004717:Endometrium / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004958:Estradiol / Q000097:blood; D005091:Exons; D005260:Female; D016147:Genes, Tumor Suppressor; D007150:Immunohistochemistry; D016912:Levonorgestrel / Q000494:pharmacology*; D008252:Macaca fascicularis; D008597:Menstrual Cycle; D015735:Mifepristone / Q000494:pharmacology*; D008938:Mitosis / Q000187:drug effects; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000097:blood; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D012333:RNA, Messenger / Q000096:biosynthesis; D011960:Receptors, Estrogen / Q000096:biosynthesis; D011980:Receptors, Progesterone / Q000096:biosynthesis; D014158:Transcription, Genetic / Q000187:drug effects*; D014599:Uterus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D016213:Cyclins; D017931:DNA Primers; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D015735:Mifepristone; D011374:Progesterone; D004958:Estradiol; D016912:Levonorgestrel