Introduction: The effect of chemotherapy treatment on premenopausal womens fertility has long been of concern. Here, we investigate the direct actions of chemotherapy drugs on the human ovary, examining effects of two commonly used drugs, cisplatin and doxorubicin.
Method: Ovarian cortical tissue samples were collected from patients aged 2734 who were undergoing elective caesarean section, with informed consent and Ethical Committee approval. Tissue was cut into small (0.5 mm×1 mm×1 mm) fragments and each piece placed on floating polycarbonate membranes in McCoys culture medium with supplements. After 24 h, cisplatin or doxorubicin were added at single-dose (5 and 1 μg/ml respectively) or double-dose (10 and 2 μg/ml respectively), or with a combination of single-doses of both drugs. Twenty-four hours later, all ovaries were moved to drug-free medium for 96 h, with bromodeoxyuridine (BrdU) added during the final 24 h of culture. Follicle number and health status was assessed histologically and levels of stroma cell proliferation (immunohistochemistry for BrdU, Abcam, Ab6326) and apoptosis (cleaved caspase-3, Cell signalling Technology, 9661S) examined.
Results and discussion: Double-doses of either cisplatin or doxorubicin resulted in a significantly greater percentage of unhealthy follicles, rising from around 30% to 6080% of all follicles (P<0.05 for both), and found specifically at the primary/secondary follicle stage (P<0.05 for both). There was no effect of single-dose of either drug. Importantly, the combination treatment, when both drugs were administered together at single-dose, also had no significant effect on follicle health. Cleaved caspase 3 expression significantly increased in the double-dose doxorubicin group and in the combination group (P<0.05 for both), while BrdU expression decreased markedly in all treatment groups apart from single-dose cisplatin (P<0.05). As such, the most marked effect of treatment was on proliferation rates, which decreased in response to the single-dose of doxorubicin, double-dose of either drug and to the combination treatment.
11 Jul 2016 - 11 Jul 2016