hsa-miR-199a-3p Inhibits Motility, Invasiveness, and Contractility of Ovarian Endometriotic Stromal Cells
hsa-miR-199a-3p Inhibits Motility, Invasiveness, and Contractility of Ovarian Endometriotic Stromal Cells
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DOI：
10.1007/s43032-021-00604-4
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发表时间：
2021-05-13
影响因子：
2.9
通讯作者：
Narahara, Hisashi
中科院分区：
文献类型：
--
作者：
Zhu, Ruofei;Nasu, Kaei;Narahara, Hisashi
提示microRNAs的异常表达参与了子宫内膜异位症的发病机制。我们先前的研究表明，microRNA hsa-miR-199 a-3 p的表达在人类增生性囊肿基质细胞（ECSC）中减弱。本研究旨在明确hsa-miR-199 a-3 p在子宫内膜异位症发生发展中的作用。分别从卵巢恶性肿瘤和正常子宫内膜组织中分离ECSC和正常子宫内膜基质细胞（NESC）。我们使用Transwell迁移测定、体外伤口愈合测定、Transwell侵袭测定和胶原凝胶收缩测定来评估转染的hsa-miR-199 a-3 p对ECSC的迁移、侵袭和收缩性的影响。我们还通过在线公共数据库搜索和荧光素酶报告基因测定检查了hsa-miR-199 a-3 p的下游靶标。hsa-miR-199 a-3 p在ECSC中的表达显著低于NESC，而p21激活激酶4（PAK 4）mRNA的表达显著高于NESC。转染hsa-miR-199 a-3 p可通过抑制PAK 4 mRNA表达抑制ECSCs的迁移、侵袭和收缩能力。PAK 4被证实是hsa-miR-199 a-3 p的直接靶点。PAK 4小干扰RNA和PAK 4抑制剂PF-3758309的转染也抑制ECSC的迁移、侵袭和收缩。这些发现表明hsa-miR-199 a-3 p可能在子宫内膜异位症的发展中起肿瘤抑制剂的作用。hsa-miR-199 a-3 p表达的减弱有利于ECSC获得增生细胞的高度侵袭性、运动性和收缩性特征，并参与子宫内膜异位症的发展。因此，PAK 4抑制剂可能有希望用于治疗子宫内膜异位症。
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.