POU2F1通过激活LDHA转录促进垂体腺瘤的恶性表型和有氧糖酵解。

POU2F1 facilitates the malignant phenotypes and aerobic glycolysis of pituitary adenoma by activating LDHA transcription.

作者信息

Wang Huanhuan, Zhang Fengjiao, Li Lei, Kang Zhiqiang, Li Jing, Mao Yu, Liu Kai, Song Lige, Shan Shuai

机构信息

Department of Endocrinology, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou City 450007 Henan, China.

出版信息

Brain Res. 2025 Aug 15;1866:149870. doi: 10.1016/j.brainres.2025.149870.

DOI:10.1016/j.brainres.2025.149870

PMID:40819704

Abstract

BACKGROUND

Pituitary adenoma (PA) can be benign, invasive, and metastatic, and its invasiveness and metastasis are poor prognosis factors for PA patients. Lactate dehydrogenase A (LDHA) plays a key role in glycolysis, and its overexpression can promote PA cell proliferation and invasion; however, the underlying mechanisms remain poorly understood.

METHODS

Reverse-transcription quantitative polymerase chain reaction and western blot were used to quantify the expression of target genes. MTT, EdU, and flow cytometry assays were utilized to detect the viability, proliferation, and apoptotic rates of HP75 cells, respectively. The invasion and migration of HP75 cells were measured by invasion and wound healing assays, and the sphere formation ability of HP75 cells was analyzed by sphere formation assay. The aerobic glycolysis of HP75 cells was tested by the corresponding kits. The JASPAR, GeneCards, and PROMO databases were used to predict the potential transcription factors of LDHA. The interaction between POU domain class 2 transcription factor 1 (POU2F1) and LDHA was verified by dual-luciferase reporter and chromatin immunoprecipitation assays. A xenograft tumor model was constructed to measure the effects of the POU2F1/LDHA axis on tumor growth in vivo.

RESULTS

LDHA was upregulated in PA tissues, and its expression levels in invasive PA tissues were higher than those in non-invasive ones. LDHA knockdown inhibited the proliferation, invasion, migration, sphere formation and aerobic glycolysis of HP75 cells and induced cell apoptosis. Mechanistically, POU2F1 activated LDHA expression to promote the proliferation, invasion, migration, sphere formation and aerobic glycolysis of HP75 cells and to inhibit cell apoptosis. Besides, LDHA overexpression could ameliorate the PI3K/AKT pathway inactivation induced by POU2F1 silence in vivo and in vitro. LDHA overexpression also alleviated the POU2F1 knockdown-induced inhibitory effect on tumor growth in vivo.

CONCLUSION

POU2F1-LDHA axis is up-regulated in pituitary adenomas and represents a potential therapeutic target for this disease.

摘要

背景

垂体腺瘤（PA）可为良性、侵袭性和转移性，其侵袭性和转移性是PA患者预后不良的因素。乳酸脱氢酶A（LDHA）在糖酵解中起关键作用，其过表达可促进PA细胞增殖和侵袭；然而，其潜在机制仍知之甚少。

方法

采用逆转录定量聚合酶链反应和蛋白质印迹法对靶基因表达进行定量。分别采用MTT、EdU和流式细胞术检测HP75细胞的活力、增殖和凋亡率。通过侵袭和伤口愈合试验检测HP75细胞的侵袭和迁移能力，并通过成球试验分析HP75细胞的成球能力。用相应试剂盒检测HP75细胞的有氧糖酵解。利用JASPAR、GeneCards和PROMO数据库预测LDHA的潜在转录因子。通过双荧光素酶报告基因和染色质免疫沉淀试验验证2类POU结构域转录因子1（POU2F1）与LDHA之间的相互作用。构建异种移植瘤模型以检测POU2F1/LDHA轴对体内肿瘤生长的影响。

结果

LDHA在PA组织中上调，其在侵袭性PA组织中的表达水平高于非侵袭性PA组织。敲低LDHA可抑制HP75细胞的增殖、侵袭、迁移、成球能力和有氧糖酵解，并诱导细胞凋亡。机制上，POU2F1激活LDHA表达，促进HP75细胞的增殖、侵袭、迁移、成球能力和有氧糖酵解，并抑制细胞凋亡。此外，LDHA过表达可在体内外改善POU2F1沉默诱导的PI3K/AKT通路失活。LDHA过表达还减轻了POU2F1敲低对体内肿瘤生长的抑制作用。