Department of Obstetrics and Gynecology, Changhai Hospital, Naval Military Medical University, No.168, Changhai Road, Yangpu District, Shanghai, 200433, China.
In Vitro Cell Dev Biol Anim. 2020 Mar;56(3):213-221. doi: 10.1007/s11626-020-00439-4. Epub 2020 Mar 17.
The proliferation and differentiation of granulosa cells are very important for follicular development. The dysfunction of granulosa cells leading to follicular development is an important cause of ovarian endocrine abnormalities. More and more evidence shows that microRNAs are involved in the regulation of ovarian granulosa cell function. It has been found that MiR-26b may be involved in CDDP resistance. Studies have shown that miR-26b can promote apoptosis of ovarian granulosa cells, but there are few studies on its mechanism, and no studies have been found on the damage of miR-26b-5p to rat ovarian granulosa cells induced by CDDP. Identification of ovarian granulosa cells was conducted by immunochemical staining. Cell counting kit 8 (CCK-8) was used to detect cell viability, flow cytometry was used to detect cell apoptosis, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot (WB) were used to analyze the expression of miR-26b-5p, MAP3K9, cleaved Caspase-3, Bax, and Bcl-2; dual-luciferase reporter assay results further verify the targeting relation between miR-26b-5p and MAP3K9. CDDP remarkably inhibited ovarian granulosa cell viability and induced ovarian granulosa cell apoptosis; miR-26b-5p inhibitor enhanced viability and inhibited apoptosis of ovarian granulosa cells, which treated with CDDP, but had little effect on normal cells. MAP3K9 partially reversed the effect of miR-26b-5p on ovarian granulosa cells induced by CDDP. miR-26b-5p has a protective effect on CDDP-induced ovarian granulosa cells via targeting MAP3K9.
颗粒细胞的增殖和分化对卵泡发育非常重要。颗粒细胞功能障碍导致卵泡发育异常是卵巢内分泌异常的重要原因。越来越多的证据表明，microRNAs 参与了卵巢颗粒细胞功能的调节。已经发现 miR-26b 可能参与 CDDP 耐药。研究表明，miR-26b 可以促进卵巢颗粒细胞凋亡，但关于其机制的研究较少，尚未发现 miR-26b-5p 对 CDDP 诱导的大鼠卵巢颗粒细胞的损伤。通过免疫化学染色鉴定卵巢颗粒细胞。使用细胞计数试剂盒 8（CCK-8）检测细胞活力，使用流式细胞术检测细胞凋亡，使用定量逆转录聚合酶链反应（qRT-PCR）和 Western blot（WB）分析 miR-26b-5p、MAP3K9、裂解 Caspase-3、Bax 和 Bcl-2 的表达；双荧光素酶报告基因实验结果进一步验证了 miR-26b-5p 与 MAP3K9 之间的靶向关系。CDDP 显著抑制卵巢颗粒细胞活力并诱导卵巢颗粒细胞凋亡；miR-26b-5p 抑制剂增强了 CDDP 处理的卵巢颗粒细胞的活力并抑制了其凋亡，但对正常细胞几乎没有影响。MAP3K9 部分逆转了 miR-26b-5p 对 CDDP 诱导的卵巢颗粒细胞的作用。miR-26b-5p 通过靶向 MAP3K9 对 CDDP 诱导的卵巢颗粒细胞具有保护作用。
