Zhang Yiming, Yu Yong, Yuan Lei, Zhang Baozhong
Department of Neurosurgery, Beijing Fengtai You'anmen Hospital, Beijing, 100069, China.
Epilepsy Center, Beijing Fengtai You'anmen Hospital, Beijing, 100069, China.
Mol Neurobiol. 2024 Nov;61(11):8668-8687. doi: 10.1007/s12035-024-04080-0. Epub 2024 Apr 1.
This study investigates the role and molecular mechanism of EZH2 in glioma cell proliferation, invasion, and migration. EZH2, miR-142-3p, lncRNA KCNQ1OT1, LIN28B, and HMGB3 expressions in glioma tissues and cells were determined using qRT-PCR or Western blot, followed by CCK-8 assay detection of cell viability, Transwell detection of invasion and migration, ChIP analysis of the enrichment of EZH2 and H3K27me3 on miR-142-3p promoter, dual-luciferase reporter assay and RIP validation of the binding of miR-142-3p-KCNQ1OT1 and KCNQ1OT1-LIN28B, and actinomycin D detection of KCNQ1OT1 and HMGB3 mRNA stability. A nude mouse xenograft model and a lung metastasis model were established. EZH2, KCNQ1OT1, LIN28B, and HMGB3 were highly expressed while miR-142-3p was poorly expressed in gliomas. EZH2 silencing restrained glioma cell proliferation, invasion, and migration. EZH2 repressed miR-142-3p expression by elevating the H3K27me3 level. miR-142-3p targeted KCNQ1OT1 expression, and KCNQ1OT1 bound to LIN28B to stabilize HMGB3 mRNA, thereby promoting its protein expression. EZH2 silencing depressed tumor growth and metastasis in nude mice via the miR-142-3p/KCNQ1OT1/HMGB3 axis. In conclusion, EZH2 curbed miR-142-3p expression, thereby relieving the inhibition of KCNQ1OT1 expression by miR-142-3p, enhancing the binding of KCNQ1OT1 to LIN28B, elevating HMGB3 expression, and ultimately accelerating glioma cell proliferation, invasion, and migration.
本研究探讨EZH2在胶质瘤细胞增殖、侵袭和迁移中的作用及分子机制。采用qRT-PCR或蛋白质免疫印迹法检测胶质瘤组织和细胞中EZH2、miR-142-3p、lncRNA KCNQ1OT1、LIN28B和HMGB3的表达，随后通过CCK-8法检测细胞活力，Transwell法检测侵袭和迁移能力，ChIP分析EZH2和H3K27me3在miR-142-3p启动子上的富集情况，双荧光素酶报告基因检测法和RIP验证miR-142-3p-KCNQ1OT1和KCNQ1OT1-LIN28B的结合情况，放线菌素D检测KCNQ1OT1和HMGB3 mRNA的稳定性。建立裸鼠异种移植模型和肺转移模型。EZH2、KCNQ1OT1、LIN28B和HMGB3在胶质瘤中高表达，而miR-142-3p低表达。EZH2沉默抑制胶质瘤细胞增殖、侵袭和迁移。EZH2通过提高H3K27me3水平抑制miR-142-3p表达。miR-142-3p靶向KCNQ1OT1表达，KCNQ1OT1与LIN28B结合以稳定HMGB3 mRNA，从而促进其蛋白表达。EZH2沉默通过miR-142-3p/KCNQ1OT1/HMGB3轴抑制裸鼠肿瘤生长和转移。总之，EZH2抑制miR-142-3p表达，从而解除miR-142-3p对KCNQ1OT1表达的抑制，增强KCNQ1OT1与LIN28B的结合，提高HMGB3表达，最终加速胶质瘤细胞增殖、侵袭和迁移。
