Yang Yun, Li Dan, Sun Lu, Liu Shasha
Department of Gynecology, Tianjin Central Hospital of Gynecology Obstetrics, China.
Adv Clin Exp Med. 2025 Jul 31. doi: 10.17219/acem/202385.
Polycystic ovary syndrome (PCOS) is a complicated endocrinological disorder.
We investigated the ferroptosis-regulated role of MALAT1 and its potential modulatory mechanisms in granulosa cells (GCs).
Reverse transcripton quantitative polymerase chain reaction (RT-qPCR) was used to measure the relative expression of MALAT1/miR-155-5p/PAK2 in KGN cells after transfection. Online bioinformatic analysis was performed to predict the interactions between MALAT1/PAK2 and miR-155-5p. Dual luciferase assays were performed for relative luciferase activity in cell groups co-transfected with the pmiRGLO plasmids containing wild type (wt) or mutant type (mt) of MALAT1 (MALAT1-wt, MALAT1-mt), siRNA targeting MALAT1(si-MALAT1) miR-155-5p inhibitor or their control was transfected into KGN cells using Lipofectamine 2000. After 48 h, the transfected cells were collected for the following experiments. Cell viability and apoptosis were measured using Cell Counting Kit-8 (CCK-8) and flow cytometry. Malondialdehyde (MDA) level and reduced glutathione (GSH) / oxidized glutathione disulfide (GSSG) ratio were detected using commercial kits. Western blot was used to measure the relative protein changes in PAK2, SLC7A11 and GPX4.
Knockdown of MALAT1 decreased cell viability, increased apoptosis and ferroptosis, which was reversed by miR-155-5p inhibition. MALAT1 downregulation inhibited PAK2, while miR-155-5p inhibition activated PAK2. The increase of relative luciferase activity in cells transfected with MALAT1-wt or PAK2-wt and miR-155-5p inhibitor suggests the bindings between miR-155-5p and MALAT1 or PAK2.
This study revealed a novel ferroptosis-modulated role of MALAT1 in PCOS in vitro via interactions with miR-155-5p/PAK2. Further in vivo and clinical studies are needed to validate these in vitro findings and fully assess the therapeutic potential of MALAT1 in PCOS.
多囊卵巢综合征（PCOS）是一种复杂的内分泌紊乱疾病。
我们研究了MALAT1在颗粒细胞（GCs）中对铁死亡的调节作用及其潜在的调节机制。
采用逆转录定量聚合酶链反应（RT-qPCR）检测转染后KGN细胞中MALAT1/miR-155-5p/PAK2的相对表达。进行在线生物信息学分析以预测MALAT1/PAK2与miR-155-5p之间的相互作用。对共转染含有野生型（wt）或突变型（mt）MALAT1（MALAT1-wt、MALAT1-mt）的pmiRGLO质粒、靶向MALAT1的小干扰RNA（si-MALAT1）、miR-155-5p抑制剂或其对照的细胞组进行双荧光素酶测定以检测相对荧光素酶活性。使用Lipofectamine 2000将上述物质转染至KGN细胞。48小时后，收集转染细胞用于后续实验。使用细胞计数试剂盒-8（CCK-8）和流式细胞术检测细胞活力和凋亡情况。使用商业试剂盒检测丙二醛（MDA）水平和还原型谷胱甘肽（GSH）/氧化型谷胱甘肽二硫化物（GSSG）比值。采用蛋白质免疫印迹法检测PAK2、SLC7A11和GPX4的相对蛋白变化。
敲低MALAT1可降低细胞活力，增加细胞凋亡和铁死亡，而抑制miR-155-5p可逆转这些变化。下调MALAT1可抑制PAK2，而抑制miR-155-5p可激活PAK2。转染MALAT1-wt或PAK2-wt以及miR-155-5p抑制剂的细胞中相对荧光素酶活性增加，提示miR-155-5p与MALAT1或PAK2之间存在结合。
本研究揭示了MALAT1在体外通过与miR-155-5p/PAK2相互作用在PCOS中对铁死亡具有新的调节作用。需要进一步的体内和临床研究来验证这些体外研究结果，并全面评估MALAT1在PCOS中的治疗潜力。
