Department of Obstetrics and Gynecology, Faculty of Medicine, Oita University, Idaigaoka 1-1, Hasama-machi, Yufu-shi, Oita, 879-5593, Japan.
Division of Obstetrics and Gynecology, Support System for Community Medicine, Faculty of Medicine, Oita University, Oita, Yufu-shi, Japan.
Reprod Sci. 2021 Dec;28(12):3498-3507. doi: 10.1007/s43032-021-00604-4. Epub 2021 May 13.
It is suggested that aberrantly expressed microRNAs are involved in the pathogenesis of endometriosis. Our previous study demonstrated that expression of the microRNA hsa-miR-199a-3p is attenuated in human endometriotic cyst stromal cells (ECSCs). The current study aimed to define the roles of hsa-miR-199a-3p in the development of endometriosis. ECSCs and normal endometrial stromal cells (NESCs) were isolated from ovarian endometrioma and normal endometrial tissues, respectively. We evaluated the effect of transfected hsa-miR-199a-3p on the migration, invasion, and contractility of ECSCs using Transwell migration assays, in vitro wound healing assays, Transwell invasion assays, and collagen gel contraction assays. We also examined the downstream target of hsa-miR-199a-3p with an online public database search and luciferase reporter assay. Expression of hsa-miR-199a-3p in ECSCs was significantly lower than that in NESCs, whereas the expression of p21-activated kinase 4 (PAK4) mRNA was significantly higher. Transfection of hsa-miR-199a-3p inhibited the migration, invasion, and contractility of ECSCs via inhibition of PAK4 mRNA expression. PAK4 was confirmed to be the direct target of hsa-miR-199a-3p. Transfection of PAK4 small interfering RNA and the PAK4 inhibitor PF-3758309 also inhibited ECSC migration, invasion, and contractility. These findings suggest that hsa-miR-199a-3p may act as a tumor suppressor in endometriosis development. Attenuation of hsa-miR-199a-3p expression was favorable for ECSCs to acquire the highly invasive, motile, and contractile characteristics of endometriotic cells and is involved in the development of endometriosis. Accordingly, PAK4 inhibitors may be promising for the treatment of endometriosis.
据报道,异常表达的 microRNA 参与了子宫内膜异位症的发病机制。我们之前的研究表明,微小 RNA hsa-miR-199a-3p 在人子宫内膜异位症囊肿基质细胞(ECSCs)中的表达减弱。本研究旨在确定 hsa-miR-199a-3p 在子宫内膜异位症发展中的作用。从卵巢子宫内膜异位囊肿和正常子宫内膜组织中分离出 ECSC 和正常子宫内膜基质细胞(NESC)。我们使用 Transwell 迁移实验、体外划痕愈合实验、Transwell 侵袭实验和胶原凝胶收缩实验评估转染 hsa-miR-199a-3p 对 ECSC 迁移、侵袭和收缩的影响。我们还通过在线公共数据库搜索和荧光素酶报告基因实验检查了 hsa-miR-199a-3p 的下游靶标。ECSCs 中 hsa-miR-199a-3p 的表达明显低于 NESCs,而 p21 激活激酶 4(PAK4)mRNA 的表达明显升高。转染 hsa-miR-199a-3p 通过抑制 PAK4 mRNA 表达抑制 ECSC 的迁移、侵袭和收缩。PAK4 被确认为 hsa-miR-199a-3p 的直接靶标。PAK4 小干扰 RNA 和 PAK4 抑制剂 PF-3758309 的转染也抑制了 ECSC 的迁移、侵袭和收缩。这些发现表明,hsa-miR-199a-3p 可能在子宫内膜异位症的发展中起肿瘤抑制作用。hsa-miR-199a-3p 表达的减弱有利于 ECSC 获得子宫内膜异位症细胞的高侵袭性、迁移性和收缩性特征,并参与了子宫内膜异位症的发展。因此,PAK4 抑制剂可能是治疗子宫内膜异位症的有前途的药物。