In vitro effect of dehydroepiandrosterone sulfate on steroid receptors, aromatase, cyclooxygenase-2 expression, and steroid hormone production in preovulatory human granulosa cells.
OBJECTIVE
To examine the effect of a low concentration of DHEAS on the expression of the androgen receptor, estrogen receptor alpha and beta, progesterone receptor, aromatase, 3-beta-hydroxysteroid dehydrogenase, and cyclooxygenase-2 in human preovulatory granulosa cells, and to measure their production of steroid hormones (estrone, estradiol, progesterone, androstenedione, and testosterone).


METHODS
Analysis of cultured primary human preovulatory granulosa cells by real-time reverse-transcriptase polymerase chain reaction and assays of hormone production.


METHODS
Osaka City University Postgraduate School of Medicine.


METHODS
Preovulatory granulosa cells were collected from follicular fluid obtained from patients undergoing transvaginal oocyte retrieval with ultrasound guidance. The cells were cultured in the absence or presence of a low concentration of DHEAS. Real-time reverse-transcriptase polymerase chain reaction was performed to quantify the RNA expression of the investigated genes, and steroid hormone (estrone, estradiol, progesterone, androstenedione, and testosterone) levels were measured in the culture medium.


METHODS
Changes in [1] the levels of mRNAs encoding androgen receptor, estrogen receptor alpha, estrogen receptor beta, progesterone receptor, aromatase, 3-beta-hydroxysteroid dehydrogenase, and cyclooxygenase-2; and [2] the levels of steroid hormones (estrone, estradiol, progesterone, androstenedione, and testosterone) in the culture medium.


RESULTS
Treatment of granulosa cells with 20 ng/mL DHEAS increased the expression of androgen receptor, aromatase, 3-beta-hydroxysteroid dehydrogenase, and cyclooxygenase-2, reduced the expression of estrogen receptor beta, and increased estrone and estradiol levels, but had no effect on progesterone, androstenedione, or testosterone levels.


CONCLUSIONS
DHEAS may be an essential trigger of ovulation.
ElBeltagy
Khalid
K
Department of Obstetrics and Gynecology, Osaka City University Graduate School of Medicine, Osaka, Japan. albeltagykak3@yahoo.com
Honda
Ken-ichi
K
Ozaki
Koji
K
Misugi
Takuya
T
Tokuyama
Osamu
O
Kimura
Masatsugu
M
Kira
Yukimi
Y
Ishiko
Osamu
O
D000328:Adult; D001141:Aromatase / Q000096:biosynthesis* / Q000235:genetics; D051546:Cyclooxygenase 2 / Q000096:biosynthesis* / Q000235:genetics; D019314:Dehydroepiandrosterone Sulfate / Q000494:pharmacology*; D005260:Female; D005498:Follicular Phase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D012739:Gonadal Steroid Hormones / Q000096:biosynthesis* / Q000235:genetics; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D011987:Receptors, Steroid / Q000096:biosynthesis* / Q000235:genetics
D012739:Gonadal Steroid Hormones; D011987:Receptors, Steroid; D019314:Dehydroepiandrosterone Sulfate; D001141:Aromatase; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human