YTHDF2介导m6A去甲基化酶ALKBH5表观沉默PLXNB1促进胶质母细胞瘤侵袭与复发的分子机制研究

胶质母细胞瘤（GBM）高度增殖与侵袭能力是其复发的主要原因，但机制不明。课题组前期：1）通过分析CGGA数据库中复发胶质瘤发现m6A去甲基化酶ALKBH5差异高表达，并证实其通过PLXNB1促进细胞增殖与迁移侵袭；2）初步证实m6A修饰识别蛋白YTHDF2在复发胶质瘤中差异高表达，且与ALKBH5表达量正相关并可负性调控PLXNB1表达，提示本研究假说：YTHDF2介导ALKBH5调控GBM细胞增殖与迁移侵袭。本研究拟从细胞、动物及临床组织标本三层面：通过体内外细胞功能学分析，明确ALKBH5促进细胞增殖与迁移侵袭；共转染证实YTHDF2介导ALKBH5表观沉默PLXNB1表达；应用MeRIP、缺失突变等技术从RNA表观遗传维度解析YTHDF2介导ALKBH5调控PLXNB1促进GBM细胞增殖与迁移侵袭的分子机制，以期为GBM侵袭与复发机制阐明及挖掘临床复发的生物标志物提供新的理论依据。

The high proliferation and invasive capacity of glioblastoma (GBM) is the main cause of recurrence, but the mechanism is unknown. Previous findings：1) High expression of m6A demethylase ALKBH5 was found by analyzing the recurrent glioma in CGGA database, and it was confirmed that it promoted cell proliferation, migration and invasion through PLXNB1. 2) Preliminary results confirmed that the m6A modified recognition protein YTHDF2 was also differentially hyperexpression in recurrent glioma, and it was positively correlated with ALKBH5 expression and negatively regulated PLXNB1 expression, suggesting that YTHDF2 mediates ALKBH5 to regulate GBM cell proliferation and migration. This study was conducted from cell, animal and clinical tissue: To identify whether ALKBH5 promotes cell proliferation, migration and invasion through in vivo and in vitro cellular functional analysis; Using co-transfection to confirm that YTHDF2 mediates ALKBH5 to epigenetic silence PLXNB1; The molecular mechanism of YTHDF2-mediated ALKBH5 regulation of PLXNB1 promoting GBM cell proliferation and migration invasion was analyzed from RNA epigenetic dimension by applying MeRIP, deletion mutation and other techniques, aim to provide a new theoretical basis for elucidating the mechanism of GBM invasion and recurrence and excavating the biomarkers of clinical recurrence.