Impaired steroidogenesis and apoptosis of granulosa-luteal cells in primary culture induced by cis-platinum.
OBJECTIVE
The purpose of this study was to test the hypothesis that the cytotoxic drug cis-platinum (CP) induces premature ovarian failure by reducing the viability of human granulosa cells.


METHODS
We incubated cultured human granulosa-luteal cells (GLCs) with varying concentrations of CP for 48 hours. Steroidogenesis and apoptosis were assessed by progesterone and estradiol, annexin V/propidium iodide, phase contrast, and transmission electron microscopy.


RESULTS
CP caused impaired production of progesterone and estradiol in a dose- and a time-dependent fashion. The estradiol production was more pronounced than progesterone for each concentration of CP that was studied. The phase contrast microscopy of CP-treated GLCs showed loss of cell number with condensed nuclei. CP-induced apoptosis was maximum at 20 μg/mL compared with a 10-μg/mL concentration (79.9% ± 4.6% vs 58.3% ± 3.9%; P < .01). The hallmark of apoptosis (ie, nuclear condensation, cell size shrinkage) was seen in CP-treated cells by transmission electron microscopy.


CONCLUSIONS
CP induces apoptosis of human GLCs in culture with impaired steroidogenesis, which may be one mechanism by which a CP-containing regime induces premature ovarian failure.
Department of Obstetrics and Gynecology, Institute for Women's Health, University College London, London, England, UK.
Ratna
R
Chatterjee
Department of Obstetrics and Gynecology, Institute for Women's Health, University College London, London, England, UK.
Mohammed
M
Helal
Department of Histopathology and Electron Microscopy, Imperial College School of Medicine, Charing Cross Hospital, London, England, UK.
Margaret
M
Mobberley
Department of Histopathology and Electron Microscopy, Imperial College School of Medicine, Charing Cross Hospital, London, England, UK.
Timothy
T
Ryder
Department of Obstetrics and Gynecology, Institute for Women's Health, University College London, London, England, UK.
Rekha
R
Bajoria
D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D002945:Cisplatin / Q000494:pharmacology*; D004958:Estradiol / Q000096:biosynthesis*; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008184:Luteal Cells / Q000187:drug effects* / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis*
D011374:Progesterone; D004958:Estradiol; D002945:Cisplatin
apoptosis; cytotoxic drug; granulosa-luteal cell; premature ovarian failure; steroidogenesis