Hedgehog信号通路介导下FOXP2对糖酵解参与子宫肌瘤细胞增殖和转移的影响

doi:10.3969/j.issn.1006-7795.2025.01.018

摘要/Abstract

摘要： 目的探讨叉头盒蛋白p2（forkhead box protein p2，FOXP2）通过调节刺猬信号通路（Hedgehog）介导的糖酵解参与子宫肌瘤细胞增殖和转移。方法从在贵州省职工医院妇科进行子宫肌瘤剔除术的10例患者中获取新鲜子宫肌瘤组织，并从中分离子宫肌瘤细胞。采用针对FOXP2的慢病毒shRNA（sh-FOXP2）以及阴性对照（sh-NC）和FOXP2过表达载体（oe-FOXP2）以及阴性对照（oe-NC）转染细胞。通过集落形成试验和Transwell检测分析评估细胞增殖、转移能力。通过葡萄糖摄取、乳酸生成的测量和细胞外酸化速率（extracellular acidification rate，ECAR）的测量检测细胞有氧糖酵解能力。使用FOXP2上调子宫肌瘤细胞建立了异种移植肿瘤模型探索FOXP2在体内的调节作用。结果与oe-NC组相比，oe-FOXP2组子宫肌瘤细胞活力、集落数量、转移细胞数、葡萄糖摄取、乳酸产生和ECAR均显著下调（P＜0.05）；与sh-NC组相比，sh-FOXP2#1组和sh-FOXP2#2组子宫肌瘤细胞活力、集落数量、转移细胞数、葡萄糖摄取、乳酸产生和ECAR均显著上调（P＜0.05）。与oe-NC+vector组相比，oe-FOXP2+vector组子宫肌瘤细胞形成的集落数量、转移细胞数、葡萄糖摄取、乳酸产生、ECAR和音猬因子（sonic hedgehog，Shh）、胶质瘤相关癌基因同源物1（glioma-associated oncogene homolog 1，Gli）蛋白均显著减少（P＜0.05）；与oe-FOXP2+vector组相比，oe-FOXP2+SAG组子宫肌瘤细胞形成的集落数量、转移细胞数、葡萄糖摄取、乳酸产生、ECAR和Shh、Gli蛋白均显著上调（P＜0.05）。与oe-NC组相比，oe-FOXP2组肿瘤质量、肿瘤增殖抗原（Ki-67）阳性细胞比例和异种移植瘤中Shh、Gli、葡萄糖转运蛋白1（glucose transporter 1，GLUT1）、磷酸甘油酸变位酶1（phosphoglycerate mutase 1，PGAM1）蛋白明显减少（P＜0.05）；与oe-FOXP2组相比，oe-FOXP2+SAG组肿瘤质量、Ki-67阳性细胞比例和异种移植瘤中Shh、Gli、GLUT1、PGAM1蛋白明显增加（P＜0.05）。结论 FOXP2通过调节Hedgehog信号通路介导的糖酵解参与子宫肌瘤细胞的增殖和转移。

关键词: 叉头盒蛋白p2, 刺猬信号通路, 糖酵解, 子宫肌瘤

Abstract: Objective To explore the role of forkhead box protein p2 (FOXP2) in the proliferation and metastasis of uterine fibroid cells by regulating glycolysis mediated by Hedgehog signaling pathway. Methods Fresh hysteromyoma tissue was obtained from 10 patients who underwent myomectomy in the Department of Gynecology of Guizhou Provincial Hospital of Workers, and hysteromyoma cells were digested and separated. Cells were transfected with lentivirus shRNA targeting FOXP2 (sh-FOXP2), negative control (sh-NC) and FOXP2 overexpression vectors (oe-FOXP2), and negative control (oe-NC). Colony formation test and Transwell assay were used to evaluate the cell proliferation and metastasis ability. Glucose uptake, lactic acid production and extracellular acidification rate (ECAR) were measured to detect the aerobic glycolysis ability of cells. A xenograft tumor model was established by up-regulating uterine leiomyoma cells with FOXP2 to explore the regulatory effect of FOXP2 in vivo. Results Compared with oe-NC group, the cell viability, colony number, migrating cell number, glucose uptake, lactic acid production and ECAR in oe-FOXP2 group decreased significantly (P<0.05). Compared with sh-NC group, the cell viability, colony number, migrating cell number, glucose uptake, lactic acid production and ECAR of uterine fibroids in sh-FOXP2#1 and sh-FOXP2#2 groups increased significantly (P<0.05). Compared with oe-NC+vector group, the number of colonies, migrating cells, glucose uptake, lactic acid production, ECAR, Shh and Gli proteins in oe-FOXP2+vector group decreased significantly (P<0.05). Compared with oe-FOXP2+vector group, the number of colonies, migrating cells, glucose uptake, lactic acid production, ECAR, Shh and Gli proteins in oe-FOXP2+SAG group were significantly increased (P<0.05). Compared with oe-NC group, the tumor weight, the proportion of Ki-67 positive cells and the proteins of Shh, Gli, GLUT1 and PGAM1 in oe-FOXP2 group decreased significantly (P<0.05). Compared with oe-FOXP2 group, the tumor weight, the proportion of Ki-67 positive cells and the proteins of Shh, Gli, GLUT1 and PGAM1 in oe-FOXP2+SAG group increased significantly (P<0.05). Conclusions FOXP2 participates in the proliferation and metastasis of uterine leiomyoma cells by regulating glycolysis mediated by Hedgehog signaling pathway.

Key words: forkhead box protein p2, Hedgehog signal pathway, glycolysis, uterine fibroids

中图分类号:

杨艳, 田妮, 龙灵, 贾振香. Hedgehog信号通路介导下FOXP2对糖酵解参与子宫肌瘤细胞增殖和转移的影响[J]. 首都医科大学学报, 2025, 46(1): 115-124.

Yang Yan, Tian Ni, Long Ling, Jia Zhenxiang. The impact of FOXP2 mediated by the Hedgehog signaling pathway on glycolysis participating in the proliferation and metastasis in uterine leiomyoma cells[J]. Journal of Capital Medical University, 2025, 46(1): 115-124.

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