miR-205-5p启动子甲基化对子宫内膜异位症患者子宫内膜细胞功能调控的研究  

Regulatory effect of miR-205-5p promoter methylation on endometrial stromal cell function in patients with endometriosis

作　　者：马菘 蔡朝霞 马颖[1] MA Song;CAI Zhaoxia;MA Ying(Department of Obstetrics and Gynecology,Zhujiang Hospital,Southern Medical University,Guangzhou 510260,China;Department of Obstetrics and Gynecology,Liwan District Maternal and Child Health Hospital,Guangzhou 510375,China)

机构地区：[1]南方医科大学珠江医院妇产医学中心,广东广州510260 [2]广州市荔湾区妇幼保健院妇产科,广东广州510375

出　　处：《新医学》2025年第1期44-52,共9页Journal of New Medicine

基　　金：广东省自然科学基金(2023A1515011688,2022A1515011880);南方医科大学珠江医院院长基金(yzjj2022ms18)。

摘　　要：目的探讨miR-205-5p启动子甲基化对子宫内膜异位症(EMS)患者子宫内膜细胞功能的调控作用。方法选择2022年10月至2023年12月在南方医科大学珠江医院行手术治疗且经病理证实的子宫内膜组织标本,根据术中及病理结果分为不存在EMS病灶的对照组(n=9)和存在EMS病灶的EMS组(n=9)。提取和分离原代子宫内膜基质细胞(ESC),采用甲基化特异性聚合酶链式反应(MSP)检测细胞miR-205-5p启动子甲基化水平,使用实时聚合酶链式反应(RT-PCR)检测去甲基化处理对miR-205-5p表达的影响,分别进行划痕-愈合、细胞增殖、细胞凋亡以及细胞侵袭实验检测去甲基化对功能的调控。将人子宫内膜碎屑埋植入皮下建立EMS小鼠模型,分为去甲基化处理组和对照组,分别于腹腔注射去甲基化制剂或生理盐水,观察2组小鼠皮下移植病灶体积及miR-205-5p表达情况。结果正常ESC启动子呈非甲基化。EMS组ESC启动子呈甲基化或高甲基化,并且使用去甲基化药物处理后,ESC的miR-205-5p mRNA表达水平升高,细胞迁移能力、增殖能力、侵袭能力减弱,细胞凋亡能力增强(均P<0.05)。经去甲基化处理后的小鼠较对照组小鼠的皮下移植病灶体积缩小,miR-205-5p表达阳性率升高(均P<0.05)。结论MiR-205-5p启动子甲基化与ESC功能调控有关,可能参与了EMS发生、发展的过程。Objective To investigate the regulatory effect of miR-205-5p promoter methylation on endometrial stromal cell(ESC)function in patients with endometriosis(EMS).Methods Endometrial tissue samples were collected from patients pathologically diagnosed with EMS undergoing surgery in Zhujiang Hospital of Southern Medical University from October 2022 to December 2023.According to intraoperative and pathological results,all samples were divided into the control group(n=9,without EMS lesions)and EMS group(n=9,with EMS lesions).Primary ESCs were extracted and isolated.Methylation-specific polymerase chain reaction(MSP)was used to detect the methylation level of miR-205-5p promoter.RTPCR was used to evaluate the effect of demethylation on the expression level of miR-205-5p.The regulatory effect of demethylation on ESC function was detected by scratch wound healing,cell proliferation,cell apoptosis and Transwell assays,respectively.EMS mouse models were established by subcutaneous seeding of human endometrial debris.All mouse models were divided into the demethylation and control groups.Demethylating agents and normal saline were injected into abdominal cavity in these two groups,respectively.The lesion size and the expression level of miR-205-5p were observed in two groups.Results Normal ESC prompter was unmethylated.In the EMS samples,the miR-205-5p promoter was methylated or hypomethylated.After demethylation,the expression level of miR-205-5p in the ESC samples was significantly upregulated,the capability of cell migration,proliferation and invasion was significantly weakened,while the ability of cell apoptosis was significantly enhanced(all P<0.05).Compared with the control group,the lesion size was decreased and the positive rate of miR-205-5p expression was increased in the demethylation group(both P<0.05).Conclusion MiR-205-5p promoter methylation is related to the regulation of ESC function in patients with EMS,which is probably involved with the incidence and progression of EMS.

关 键 词：子宫内膜异位症 miR-205-5p 甲基化 子宫内膜细胞 

分 类 号：R73[医药卫生—肿瘤]