Notch1信号通过调控细胞增殖、迁移、侵袭和糖酵解参与子宫腺肌病的发生及发展
Role of Notch 1 signaling and glycolysis in the pathogenic mechanism of adenomyosis
温小慧 1黄诗雅 1刘学红 1李坤寅 2关永格2
作者信息
- 1. 广州中医药大学第三临床医学院,广东 广州 510400;广州中医药大学岭南医学研究中心,广东 广州 510400
- 2. 广州中医药大学第三附属医院妇科,广东 广州 510100
- 折叠
摘要
目的 基于Notch1信号观察子宫腺肌病(AM)在位内膜及Ishikawa细胞系糖酵解相关因子和蛋白的表达,并探讨子宫腺肌病的发生机制.方法 收集AM(AM组,n=8)及子宫肌瘤(CON组,n=8)在位子宫内膜组织,通过临床特征评估两组子宫内膜的可比性,qRT-PCR和Western blot检测其Notch1信号及糖酵解相关因子和蛋白的表达,通过葡萄糖和乳酸试验检测其葡萄糖和乳酸的含量;建立过表达Notch1的Ishikawa细胞慢病毒稳转株,CCK-8检测其细胞存活率,Transwell迁移和侵袭实验检测其迁移和侵袭能力,细胞外酸化速率检测其糖酵解储备.结果 与CON组子宫内膜组织比较,AM组糖类抗原125(CA125)水平明显高于CON组(P<0.01),AM患者在位子宫内膜组织Notch1、HK2和PDHA的mRNA相对表达量增加(P<0.01或P<0.05),Notch1、GLUT1、HK2、PKM和PDHA蛋白相对表达量均显著升高(P<0.01或P<0.05),葡萄糖含量显著下降、乳酸水平显著升高(P<0.01或P<0.05).与对照病毒空载体(ov-NC)比较,过表达Notch1的Ishikawa细胞慢病毒稳转株(ov-Notch1)的细胞存活率显著升高(P<0.05),细胞迁移和侵袭细胞数量显著增加(P<0.05),酵解容量和酵解储备显著升高(P<0.01或P<0.05).结论 Notch1信号可能通过调控细胞的增殖、迁移、侵袭和糖酵解过程,进而参与AM的发生发展.
Abstract
Objective To investigate the expressions of glycolysis-related factors and changes in Notch1 signaling in endometrial tissues of adenomyosis(AM)and Ishikawa cells to explore the pathogenesis of AM.Methods Eutopic endometrial tissues were collected from 8 patients with AM and 8 patients with uterine fibroids matched for clinical characteristics(control group).The expressions of Notch1 signaling proteins and glycolysis-related factors in the collected tissues were detected using qRT-PCR and Western blotting,and the levels of glucose and lactic acid were determined.An Ishikawa cell model with lentivirus-mediated stable Notch1 overexpression was established for assessing cell survival rate with CCK-8 assay,cell migration and invasion abilities with Transwell migration and invasion assays,and glycolytic capacity by determining the extracellular acidification rate.Results Compared with those in the control group,the endometrial tissues in AM group showed significantly increased expression level of carbohydrate antigen 125(CA125),increased mRNA expression levels of Notch1,HK2 and PDHA and protein expressions of Notch1,GLUT1,HK2,PKM and PDHA,lowered glucose level and increased lactate level.The Ishikawa cell models with stable Notch1 overexpression exhibited significantly increased cell survival rate with attenuated cell migration and invasion abilities and decreased glycolysis capacity and reserve.Conclusion The Notch1 signaling pathway participates in the pathogenesis of AM possibly by regulating the proliferation,migration,invasion and glycolysis of endometrial cells.
关键词
子宫腺肌病/子宫内膜/糖酵解/Ishikawa细胞系/慢病毒Key words
adenomyosis/endometrium/glycolysis/Ishikawa/lentivirus引用本文复制引用
基金项目
广东省普通高校重点领域专项项目(2023ZDZX2011)
广东省普通高校重点领域专项项目(2021ZDZX2015)
广州中医药大学"双一流"与高水平大学学科协同创新团队项目()
广州中医药大学中医学学科中医基础理论研究"揭榜挂帅"项目()
出版年
2024
国家科技期刊平台
NSTL
万方数据