摘要
目的:探讨Enolase 1(ENO1)调节急性髓系白血病(AML)细胞的代谢及分子机制。方法:分析GEO、TCGA等数据库中AML患者ENO1表达情况;运用Kaplan-Meier分析基因表达水平与患者生存期之间的关系;构建ENO1 shRNA慢病毒质粒体系,感染MV4-11、NB4细胞系构建分组,应用Western blot法检测ENO1在AML两种细胞系中的敲降效率;CCK-8法检测其对AML细胞生长的影响;Annexin V-PE/7-AAD法检测细胞凋亡情况;应用GO、KEGG富集方法分析ENO1在细胞中的功能及相关信号通路。结果:通过GEO、TCGA数据库分析表明ENO1在多数AML患者体内显著高表达(P<0.05,P<0.01),而在其它类型白血病患者中无此变化;ENO1在AML细胞中,在白血病干细胞,在AML小鼠模型造血干细胞中均显著高表达(P<0.001)。Kaplan-Meier分析表明ENO1高表达AML患者生存期显著低于ENO1低表达患者(P<0.05,HR>1)。敲降ENO1显著抑制AML细胞生长,诱导AML细胞凋亡(P<0.01)。富集分析发现,AML细胞中ENO1可能参与组蛋白甲基化、RNA干扰复合物的形成、糖代谢相关生物过程;其负相关共表达基因主要富集在自噬和FOXO信号通路。结论:ENO1在AML患者中尤其是AML干细胞中高表达并与生存期呈负相关。且ENO1高表达可促进AML细胞生长、抑制AML细胞凋亡,ENO1有望为AML患者的治疗和预后提供一定的参考。
Objective:To investigate the role and molecular mechanism of Enolase 1(ENO1)in regulating cell metabolism in acute myeloid leukemia(AML).Methods:The expression of ENO1 in AML patients was analyzed from GEO,TCGA and other databases.The relationship between gene expression levels and patient overall survival was analyzed by applying Kaplan-Meier analysis.ENO1 shRNA lentiviral plasmid system was constructed,then infected MV4-11 and NB4 cell lines respectively.Western blot method was used to detect the knockdown efficiency of ENO1.CCK-8 method were used to detect its effect on AML cell growth.Annexin V-PE/7-AAD staining was used to explore the apoptosis on AML cell.GO and KEGG protein enrichment methods were used to analyze the function of ENO1 in cells and its related signaling pathways.Results:Analysis through GEO and TCGA databases showed that ENO1 was significantly highly expressed in most AML patients(P<0.05,P<0.01),but not in patients with other types of leukemia.ENO1 was significantly highly expressed in AML cells,in leukemic stem cells,and in hematopoietic stem cells in AML mouse models(P<0.001).Kaplan-Meier analysis showed that patients with high ENO1 expression had significantly lower overall survival than those with low ENO1 expression(P<0.05,HR>1).Knockdown of ENO1 significantly inhibited AML cell growth and induced AML cell apoptosis(P<0.01).Enrichment analysis revealed that in AML cells,ENO1 was mainly involved in histone methylation,RNA interference complex formation and glucose metabolism.Its negatively coexpression genes were principally enriched in autophagy and FOXO signaling pathway.Conclusion:ENO1 is highly expressed in AML patients,especially in AML stem cells and is negatively correlated with overall survival.Moreover,high expression of ENO1 can promote the growth of AML cells and inhibit the apoptosis of AML cells.ENO1 is expected to provide some reference for the treatment and prognosis of AML patients.
作者
宋鹏
史贺文
邹颖
李敏敬
杨春燕
SONG Peng;SHI Hewen;ZOU Ying;LI Minjing;YANG Chunyan(Institute of Integrated Medicine,Binzhou Medical University,Shandong Yantai 264003,China;Medicine&Pharmacy Research Center,Binzhou Medical University,Shandong Yantai 264003,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第24期4524-4531,共8页
Journal of Modern Oncology
基金
山东省自然科学基金(编号:ZR2019PH080,ZR2020MH120)。
