Dipartimento di Medicina Sperimentale, Cattedra di Endocrinologia, Sapienza Università di Roma, c/o Servizio Speciale Malattie della Tiroide, Viale Regina Elena, Rome, Italy.
J Endocrinol. 2012 Nov;215(2):281-9. doi: 10.1530/JOE-12-0153. Epub 2012 Aug 21.
Infertility is a dramatic and frequent side effect in women who are undergoing chemotherapy. Actual strategies are mainly focused on oocyte cryopreservation, but this is not always a suitable option. Considering the key role that granulosa cells play in follicle life, we studied whether thyroid hormone 3,5,3'-triiodothyronine (T(3)) protects rat ovarian granulosa cells from chemotherapy-induced apoptosis. To this aim, a cell line was established from fresh isolated rat granulosa cells and named rGROV. Cells were exposed to paclitaxel (PTX) and T(3), and apoptosis, cell viability, and cell cycle distribution were analyzed under different conditions. First, the integrity of the steroidogenic pathway was demonstrated, and the presence of thyroid receptors, transporters, and deiodinases was confirmed by quantitative PCR. Cells were then exposed to PTX alone or contemporary to T(3). MTT and TUNEL assays revealed that while there was a relevant percentage of dying cells when exposed to PTX (40-60%), the percentage was sensibly reduced (20-30%) in favor of living cells if T(3) was present. Cell cycle analysis showed that cells exposed to PTX alone were first collected in G2 and then died by apoptosis; on the other hand, the T(3) granted the cells to cycle regularly and survive PTX insult. In addition, western blot and FCM analyses confirmed that caspases activation, casp 3 and Bax, were downregulated by T(3) and that Bcl2 and cyclins A and B together with cdk1 were upregulated by T(3). In conclusion, we demonstrated that thyroid hormone T(3) can counteract the lethal effect of taxol on granulosa cells.
不孕是接受化疗的女性中常见且显著的副作用。目前的策略主要集中在卵母细胞冷冻保存上，但这并不总是一个合适的选择。鉴于颗粒细胞在卵泡寿命中起着关键作用，我们研究了甲状腺激素 3,5,3'-三碘甲状腺原氨酸（T3）是否可以保护大鼠卵巢颗粒细胞免受化疗诱导的凋亡。为此，我们从新鲜分离的大鼠颗粒细胞中建立了细胞系，并将其命名为 rGROV。将细胞暴露于紫杉醇（PTX）和 T3 中，并在不同条件下分析细胞凋亡、细胞活力和细胞周期分布。首先，我们证明了类固醇生成途径的完整性，并通过定量 PCR 证实了甲状腺受体、转运蛋白和脱碘酶的存在。然后，我们将细胞单独暴露于 PTX 或与 T3 同时暴露。MTT 和 TUNEL 测定表明，当暴露于 PTX 时，有相当比例的细胞死亡（40-60%），但如果存在 T3，则死亡细胞的比例明显降低（20-30%），有利于存活细胞。细胞周期分析表明，单独暴露于 PTX 的细胞首先被收集在 G2 期，然后通过凋亡死亡；另一方面，T3 使细胞能够正常循环并耐受 PTX 的损伤。此外，Western blot 和 FCM 分析证实，T3 下调了 caspase 激活、caspase 3 和 Bax，上调了 Bcl2 以及细胞周期蛋白 A 和 B 与 cdk1。总之，我们证明了甲状腺激素 T3 可以抵抗紫杉醇对颗粒细胞的致命作用。
