Xiao Chenhu, Hou Ge, Wang Cheng, Huang Yangyang, Liu Zongwen
Department of Radiation Oncology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of Radiation Oncology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Pathol Res Pract. 2025 Aug;272:156102. doi: 10.1016/j.prp.2025.156102. Epub 2025 Jun 30.
Glycolysis can contribute to radiotherapy resistance in cancers. Methyltransferase-like protein-1 (METTL1) and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) are oncogenes in esophageal cancer (EC). Herein, the interaction between METTL1 and PFKFB3 in regulating glycolysis and radioresistance in EC was explored.
PFKFB3 and METTL1 mRNA and protein levels were determined by RT-qPCR and Western blot. Cell viability, apoptosis, invasion, migration, and stemness were respectively measured via cell counting kit-8 assay, flow cytometry, transwell assay, wound healing assay and sphere formation assay. Glycolytic metabolism was evaluated by glucose consumption, lactate production, ATP/ADP ratio, and seahorse assay. Radiotherapy resistance was assessed using cell survival assay. RNA immunoprecipitation was performed to confirm interaction between METTL1 and PFKFB3. PFKFB3 function in vivo was studied using xenograft tumor model in mice.
PFKFB3 was significantly up-regulated in radiation-treated EC cells. Downregulation of PFKFB3 impeded EC cell viability, invasion, migration, stemness. PFKFB3 knockdown suppressed glycolysis to reduce radiation resistance of EC cells. METTL1 could enhance PFKFB3 mRNA stability by mediating the m7G modification of PFKFB3. METTL1 facilitated cell development, glycolysis and radioresistance via promoting PFKFB3. Radioresistance of EC was also decreased after silence of PFKFB3 in vivo.
This study evidenced that METTL1 induced m7G modification to stabilize PFKFB3 mRNA that further enhanced radiation resistance in EC via stimulating glycolysis.
糖酵解可导致癌症放疗抵抗。甲基转移酶样蛋白1（METTL1）和6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）是食管癌（EC）中的癌基因。在此，探讨了METTL1与PFKFB3在调节EC糖酵解和放疗抵抗中的相互作用。
通过RT-qPCR和蛋白质印迹法测定PFKFB3和METTL1的mRNA和蛋白质水平。分别通过细胞计数试剂盒-8检测、流式细胞术、Transwell检测、伤口愈合检测和球体形成检测来测量细胞活力、凋亡、侵袭、迁移和干性。通过葡萄糖消耗、乳酸生成、ATP/ADP比值和海马检测评估糖酵解代谢。使用细胞存活检测评估放疗抵抗。进行RNA免疫沉淀以确认METTL1与PFKFB3之间的相互作用。使用小鼠异种移植肿瘤模型研究PFKFB3在体内的功能。
PFKFB3在放疗处理的EC细胞中显著上调。PFKFB3的下调阻碍了EC细胞的活力、侵袭、迁移和干性。PFKFB3敲低抑制糖酵解以降低EC细胞的放疗抵抗。METTL1可通过介导PFKFB3的m7G修饰来增强PFKFB3 mRNA稳定性。METTL1通过促进PFKFB3促进细胞发育、糖酵解和放疗抵抗。在体内沉默PFKFB3后，EC的放疗抵抗也降低。
本研究证明，METTL1诱导m7G修饰以稳定PFKFB3 mRNA，进而通过刺激糖酵解增强EC的放疗抵抗。
