miR-130b regulates gap junctional intercellular communication through connexin 43 in granulosa cells from patients with polycystic ovary syndrome
miR-130b regulates gap junctional intercellular communication through connexin 43 in granulosa cells from patients with polycystic ovary syndrome
复制标题
miR-130b 通过连接蛋白 43 调节多囊卵巢综合征患者颗粒细胞中的间隙连接细胞间通讯
DOI：
10.1093/molehr/gaaa044
复制
发表时间：
2020-08-01
影响因子：
4
通讯作者：
Yang, Dongzi
中科院分区：
文献类型：
--
作者：
Jiang, Linlin;Huang, Hui;Yang, Dongzi
MicroRNA (miRNA) 是一种小型非编码 RNA，可在转录后负向调节基因表达。我们探讨了与正常月经周期女性的黄素化颗粒细胞相比，多囊卵巢综合征 (PCOS) 女性的黄素化颗粒细胞中连接蛋白 43 (Cx43) 的表达是否存在差异，以及某些 miRNA 是否调节 Cx43 水平和间隙连接细胞间通讯 (GJIC)。使用 miRNA 微阵列对五名患有 PCOS 的女性和五名未患 PCOS 的女性的卵巢皮质组织中的 miRNA 进行了研究。使用实时 PCR 测量 20 名患有 PCOS 的女性和 25 名未患有 PCOS 的女性的人黄素化颗粒细胞中 miR-130b 和 Cx43 mRNA 的水平。进行蛋白质和 mRNA 表达分析以及荧光素酶测定以确认 miR-130b 的底物。与非 PCOS 卵巢皮质相比，PCOS 卵巢皮质显示出 miRNA 的差异表达。此外，与正常月经周期的女性相比，PCOS 卵巢皮质和黄素化颗粒细胞中的 miR-130b 水平升高，而 PCOS 患者颗粒细胞中 miR-130b 的确定靶点 Cx43 mRNA 水平降低。使用刮片加载和染料转移测定，颗粒细胞系中 miR-130b 的过度表达导致 Cx43 蛋白水平降低并抑制 GJIC。同时，抑制 miR-130b 会增加 Cx43 水平。总之，miR-130b 在 PCOS 颗粒细胞中增加，它靶向 Cx43 来影响 GJIC。本研究结果提示miR-130b通过Cx43的转录后调控参与PCOS的病理生理学，为了解PCOS的病理机制提供了新的见解。
MicroRNAs (miRNAs) are small, noncoding RNAs that negatively regulate gene expression post-transcriptionally. We explored whether connexin 43 (Cx43) was differently expressed in luteinized granulosa cells from women with polycystic ovary syndrome (PCOS) compared with luteinized granulosa cells from women with a normal menstrual cycle, and whether certain miRNAs regulate the Cx43 level and gap junctional intercellular communication (GJIC). The miRNA profile was investigated in ovarian cortex tissues from five women with PCOS and five women without PCOS using a miRNA microarray. The levels of miR-130b and Cx43 mRNA were measured using real-time PCR in human luteinized granulosa cells from 20 women with PCOS and 25 women without PCOS. Protein and mRNA expression analysis and luciferase assays were conducted to confirm the substrate of miR-130b. PCOS ovarian cortex showed differential expression of miRNAs compared with non-PCOS ovarian cortex. Furthermore, miR-130b levels were increased in PCOS ovarian cortex and in luteinized granulosa cells compared with those in women with normal menstrual cycles, whereas the level of Cx43 mRNA, the identified target of miR-130b, was decreased in granulosa cells from patients with PCOS. Overexpression of miR-130b in a granulosa cell line resulted in reduced Cx43 protein levels and inhibited GJIC using scrape loading and dye transfer assay. Meanwhile, inhibition of miR-130b increased the Cx43 level. In conclusion, miR-130b was increased in PCOS granulosa cells, where it targets Cx43 to affect GJIC. The results of the present study suggested that miR-130b, via post-transcriptional regulation of Cx43, is involved in the pathophysiology of PCOS, which provides new insight into the pathological mechanism of PCOS.