ALKBH5介导m6A修饰调控Galectin-9促进异位内膜基质细胞侵袭、迁移与增殖的机制研究  

Mechanism of ALKBH5 mediated m6A regulation of Galectin-9 in the invasion, migration, and proliferation of endometrial stromal cell

作　　者：王芳 刘恒炜 梁嘉欣 王细文[2] WANG Fang;LIU Hengwei;LIANG Jiaxin;WANG Xiwen(Department of Gynecology and Obstetrics,Xinjiang Uygur Autonomous Region Occupational Disease Hospital,Urumqi 830052,China;Department of Gynecology,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]新疆维吾尔自治区职业病医院妇产科,乌鲁木齐830052 [2]武汉大学中南医院妇科,武汉430071

出　　处：《医学新知》2024年第5期487-496,共10页New Medicine

基　　金：国家自然科学基金青年科学基金项目(82001524);湖北省自然科学基金(2020CFB310);新疆维吾尔自治区自然科学基金(2021D01A165)。

摘　　要：目的探讨ALKBH5对Galectin-9的m6A修饰作用在子宫内膜基质细胞(endometrial stromal cells,ESCs)侵袭、迁移和增殖过程中的潜在影响。方法收集20例正常子宫内膜(normal endometrium,NC)组织及20例子宫内膜异位症患者异位子宫内膜(ectopic endometrium,EC)组织,使用实时荧光定量PCR(qRT-PCR)、Western blot及免疫组织化学法检测ALKBH5和Galectin-9在组织中的表达。同时收集正常增生期子宫内膜组织10例并提取原代ESCs,通过生物信息学分析、甲基化RNA免疫共沉淀(MeRIP)、RNA免疫共沉淀(RIP)实验探究ALKBH5调控Galectin-9 m6A水平和mRNA表达的作用机制。过表达ALKBH5后,通过Transwell、细胞计数试剂盒8(CCK-8)法测定各组ESCs细胞增殖、迁移、侵袭能力,研究ALKBH5对ESCs侵袭、迁移和增殖的影响。结果与NC组织相比,EC组织中ALKBH5和Galectin-9表达升高(P<0.05)。过表达ALKBH5能够降低Galectin-9的m6A修饰水平,增强Galectin-9 mRNA的稳定性和表达。过表达ALKBH5能够促进ESCs侵袭迁移和增殖,而使用siRNA沉默Galectin-9可有效逆转ALKBH5介导的ESCs细胞侵袭、迁移和增殖(P<0.05)。结论ALKBH5通过降低Galectin-9 m6A修饰水平,上调Galectin-9 mRNA表达,从而促进ESCs侵袭、迁移与增殖。Objective To investigate the potential role of ALKBH5-mediated m6A modification of Galectin-9 in the invasion,migration,and proliferation of endometrial stromal cells(ESCs)in endometriosis.Methods A total of 20 normal endometrial(NC)specimens and 20 ectopic endometrial(EC)specimens from patients with endometriosis were obtained.The expression of ALKBH5 and Galectin-9 in tissues was detected using immunohistochemistry,qRT-PCR and Western blot.In addition,10 cases of normal proliferative endometrial tissues were collected and primary ESCs were extracted,and the mechanism of ALKBH5 in regulating the level and mRNA expression of Galectin-9 m6A was investigated by bioinformatics analysis,methylated RNA immunoprecipitation(MeRIP),and RIP assay.After ALKBH5 overexpression,the proliferation,migration,and invasion abilities of ESCs were determined by Transwell and cell counting kit-8(CCK-8)assays.The effect of ALKBH5 on ESCs invasion,migration,and proliferation was investigated.Results Compared with NC tissues,the expression of ALKBH5 and Galectin-9 was increased in EC tissues from patients with endometriosis(P<0.05).ALKBH5 overexpression could reduce the m6A modification level of Galectin-9 and enhance the stability and expression of Galectin-9 mRNA.ALKBH5 overexpression promoted the invasion,migration,and proliferation of ESCs,while Galectin-9 knockdown partially reversed the promoting effect of ALKBH5 overexpression on ESCs invasion,migration,and proliferation(P<0.05).Conclusion ALKBH5 promotes ESCs invasion,migration,and proliferation by promoting Galectin-9 mRNA expression via reducing m6A modification.

关 键 词：子宫内膜异位症 m6A修饰 ALKBH5 GALECTIN-9 侵袭 

分 类 号：R711.71[医药卫生—妇产科学]