无功能垂体腺瘤（NFPA）是垂体腺瘤（PA）的主要亚型之一，其主要治疗方法是手术切除。然而，普通手术无法彻底清除病灶，一线治疗仍缺乏，因此针对NFPA新药的研发无疑刻不容缓。据报道，冬凌草甲素 (ORI) 对多种肿瘤具有抗肿瘤作用，但它是否对 NFPA 具有相同的作用还需要进一步研究。通过细胞计数试剂盒-8、集落形成测定、5-乙炔基-2'-脱氧尿苷增殖测定检查 ORI 对垂体来源的滤泡细胞系 (PDFS) 细胞活力、集落形成、增殖能力、迁移和侵袭的影响、伤口愈合测定和Transwell测定。通过转录组测序分析筛选对照组和ORI处理组中差异表达的基因，并通过京都基因和基因组百科全书（KEGG）和基因本体论（GO）富集进行分析。进行细胞周期分析以检测细胞周期的变化。进行膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色以检测ORI处理的细胞中的细胞凋亡。进行Western blot检测Bax、Bcl-2和cleaved Caspase-3蛋白的表达。 ORI 抑制 PDFS 细胞活力并显着抑制细胞增殖、迁移和侵袭。 GO和KEGG结果显示ORI与PDFS细胞中的细胞周期和凋亡等信号通路相关。此外，ORI 阻断 G2/M 期细胞并诱导 PDFS 细胞凋亡。 ORI 可以在 PDFS 细胞中协同触发细胞周期破坏和细胞凋亡，使其成为 NFPA 治疗有前途且有效的药物。© 2024 Wiley periodicals LLC。

Nonfunctioning pituitary adenoma (NFPA) is one of the major subtypes of pituitary adenomas (PA) and its primary treatment is surgical resection. However, normal surgery fails to remove lesions completely and there remains in lack of frontline treatment, so the development of new drugs for NFPA is no doubt urgent. Oridonin (ORI) has been reported to have antitumor effects on a variety of tumors, but whether it could exhibit the same effect on NFPA requires to be further investigated. The effects of ORI on pituitary-derived folliculostellate cell line (PDFS) cell viability, colony formation, proliferation ability, migration, and invasion were examined by Cell Counting Kit-8, colony formation assay, 5‑Ethynyl‑2'‑deoxyuridine proliferation assay, wound-healing assay, and Transwell assay. The differentially expressed genes in the control and ORI-treated groups were screened by transcriptome sequencing analysis and analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment. Cell cycle analysis was performed to detect changes in cell cycle. Annexin V-fluorescein isothiocyanate/propidium iodide staining was performed to detect apoptosis in ORI-treated cells. Western blot assay was performed to detect Bax, Bcl-2, and cleaved Caspase-3 protein expression. ORI inhibited PDFS cell viability and significantly suppressed cell proliferation, migration, and invasion. GO and KEGG results showed that ORI was associated with signaling pathways such as cell cycle and apoptosis in PDFS cells. In addition, ORI blocked cells in G2/M phase and induced apoptosis in PDFS cells. ORI can trigger cell cycle disruption and apoptosis collaboratively in PDFS cells, making it a promising and effective agent for NFPA therapy.© 2024 Wiley Periodicals LLC.