HOXC6(同源异型框C6)和甲基转移酶样3(METTL3)已显示与前列腺癌(PCa)的进展有关。然而,尚未报道HOXC6是否通过METTL3介导的N6-甲基腺苷(m6A)修饰在PCa中产生致癌作用.细胞计数套件-8(CCK-8),5-乙炔基-2'-脱氧尿苷(EdU),流式细胞术,transwell,划痕,球体形成试验用于细胞生长,入侵,迁移和干性分析。通过测量葡萄糖消耗来评估糖酵解,乳酸生成和ATP/ADP比。通过RNA免疫沉淀(Me-RIP)测定确定N6-甲基腺嘌呤(m6A)修饰谱。与PGK1(磷酸甘油酸激酶1)相互作用的蛋白质通过免疫共沉淀测定进行确认。在小鼠中进行肿瘤形成实验用于体内测定。PCa组织和细胞显示高表达HOXC6和METTL3。功能上,HOXC6或METTL3的沉默抑制PCa细胞增殖,入侵,迁移,stemness,和糖酵解。此外,METTL3诱导HOXC6m6A修饰以稳定其表达。此外,m6A阅读器IGF2BP2直接识别并结合HOXC6mRNA,并保持其稳定性,并参与METTL3对HOXC6表达的调控。此外,IGF2BP2敲低受损PCa细胞增殖,入侵,迁移,stemness,和通过调节HOXC6的糖酵解。此外,HOXC6还与PCa细胞中的糖化酶PGK1相互作用。体内测定进一步显示METTL3沉默降低HOXC6和PGK1的表达,并阻碍PCa生长。METTL3通过在m6A-IGF2BP2依赖性机制中维持HOXC6表达促进PCa进展。
HOXC6 (Homeobox C6) and methyltransferase-like 3 (METTL3) have been shown to be involved in the progression of prostate cancer (PCa). However, whether HOXC6 performs oncogenic effects in PCa via METTL3-mediated N6-methyladenosine (m6A) modification is not yet reported. The Cell Counting Kit-8 (CCK-8), 5-ethynyl-2\'-deoxyuridine (EdU), flow cytometry, transwell, scratch, sphere formation assays were applied for cell growth, invasion, migration and stemness analyses. Glycolysis was evaluated by measuring glucose consumption, lactate generation and ATP/ADP ratio. The N6-methyladenine (m6A) modification profile was determined by RNA immunoprecipitation (Me-RIP) assay. The proteins that interact with PGK1 (phosphoglycerate kinase 1) were confirmed by Co-immunoprecipitation assay. Tumor formation experiments in mice were conducted for in vivo assay. PCa tissues and cells showed highly expressed HOXC6 and METTL3. Functionally, the silencing of HOXC6 or METTL3 suppresses PCa cell proliferation, invasion, migration, stemness, and glycolysis. Moreover, METTL3-induced HOXC6 m6A modification to stabilize its expression. In addition, the m6A reader IGF2BP2 directly recognized and bound to HOXC6 mRNA, and maintained its stability, and was involved in the regulation of HOXC6 expression by METTL3. Furthermore, IGF2BP2 knockdown impaired PCa cell proliferation, invasion, migration, stemness, and glycolysis by regulating HOXC6. Besides that HOXC6 interacted with the glycoytic enzyme PGK1 in PCa cells. In vivo assays further showed that METTL3 silencing reduced the expression of HOXC6 and PGK1, and impeded PCa growth. METTL3 promoted PCa progression by maintaining HOXC6 expression in an m6A-IGF2BP2-dependent mechanism.