MALAT1 modulates granulosa cells ferroptosis and apoptosis through PAK2 upregulation in polycystic ovary syndrome.

IF 1.9 4区医学 Q3 MEDICINE, RESEARCH & EXPERIMENTAL

Advances in Clinical and Experimental Medicine Pub Date : 2025-07-31 DOI:10.17219/acem/202385

Yun Yang, Dan Li, Lu Sun, Shasha Liu

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引用次数: 0

Abstract

Background: Polycystic ovary syndrome (PCOS) is a complicated endocrinological disorder.

Objectives: We investigated the ferroptosis-regulated role of MALAT1 and its potential modulatory mechanisms in granulosa cells (GCs).

Material and methods: 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.

Results: 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.

Conclusions: 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.

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MALAT1通过上调PAK2调控多囊卵巢综合征中颗粒细胞的铁下垂和凋亡。

背景：多囊卵巢综合征（PCOS）是一种复杂的内分泌疾病。目的：研究MALAT1在颗粒细胞（GCs）中对铁凋亡的调节作用及其潜在的调节机制。材料与方法：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的相对表达量。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。用含有野生型（wt）或突变型（mt） MALAT1（MALAT1-wt, MALAT1-mt）的pmiRGLO质粒共转染的细胞组进行双荧光素酶测定，使用Lipofectamine 2000将靶向MALAT1(si-MALAT1) miR-155-5p抑制剂的siRNA或其对照转染到KGN细胞中。48 h后，收集转染后的细胞进行后续实验。采用细胞计数试剂盒-8 （CCK-8）和流式细胞术检测细胞活力和凋亡。采用商品化试剂盒检测丙二醛（MDA）水平和还原型谷胱甘肽(GSH) /氧化谷胱甘肽二硫（GSSG）比值。Western blot检测PAK2、SLC7A11和GPX4蛋白的相对变化。结果：MALAT1的敲低降低了细胞活力，增加了细胞凋亡和铁下垂，而miR-155-5p的抑制可以逆转这一现象。MALAT1下调抑制PAK2，而miR-155-5p抑制激活PAK2。在转染MALAT1-wt或PAK2-wt和miR-155-5p抑制剂的细胞中，荧光素酶的相对活性增加，表明miR-155-5p与MALAT1或PAK2结合。结论：本研究揭示了MALAT1通过与miR-155-5p/PAK2的相互作用，在体外PCOS中具有新的铁凋亡调节作用。需要进一步的体内和临床研究来验证这些体外研究结果，并充分评估MALAT1在多囊卵巢综合征中的治疗潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Advances in Clinical and Experimental Medicine MEDICINE, RESEARCH & EXPERIMENTAL-

CiteScore

3.70

自引率

4.80%

发文量

153

审稿时长

6-12 weeks

期刊介绍： Advances in Clinical and Experimental Medicine has been published by the Wroclaw Medical University since 1992. Establishing the medical journal was the idea of Prof. Bogumił Halawa, Chair of the Department of Cardiology, and was fully supported by the Rector of Wroclaw Medical University, Prof. Zbigniew Knapik. Prof. Halawa was also the first editor-in-chief, between 1992-1997. The journal, then entitled "Postępy Medycyny Klinicznej i Doświadczalnej", appeared quarterly. Prof. Leszek Paradowski was editor-in-chief from 1997-1999. In 1998 he initiated alterations in the profile and cover design of the journal which were accepted by the Editorial Board. The title was changed to Advances in Clinical and Experimental Medicine. Articles in English were welcomed. A number of outstanding representatives of medical science from Poland and abroad were invited to participate in the newly established International Editorial Staff. Prof. Antonina Harłozińska-Szmyrka was editor-in-chief in years 2000-2005, in years 2006-2007 once again prof. Leszek Paradowski and prof. Maria Podolak-Dawidziak was editor-in-chief in years 2008-2016. Since 2017 the editor-in chief is prof. Maciej Bagłaj. Since July 2005, original papers have been published only in English. Case reports are no longer accepted. The manuscripts are reviewed by two independent reviewers and a statistical reviewer, and English texts are proofread by a native speaker. The journal has been indexed in several databases: Scopus, Ulrich’sTM International Periodicals Directory, Index Copernicus and since 2007 in Thomson Reuters databases: Science Citation Index Expanded i Journal Citation Reports/Science Edition. In 2010 the journal obtained Impact Factor which is now 1.179 pts. Articles published in the journal are worth 15 points among Polish journals according to the Polish Committee for Scientific Research and 169.43 points according to the Index Copernicus. Since November 7, 2012, Advances in Clinical and Experimental Medicine has been indexed and included in National Library of Medicine’s MEDLINE database. English abstracts printed in the journal are included and searchable using PubMed http://www.ncbi.nlm.nih.gov/pubmed.