Zhang Chunhua, Liu Qing, Bi Lei, Chen Weiping, Zeng Li
School of Chinese Medicine, Macau University of Science and Technology, Macau, China.
Department of Gynecology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China.
Scand J Immunol. 2025 Apr;101(4):e70017. doi: 10.1111/sji.70017.
We aimed to investigate the effects of Salvianolic acid A (SA), an active ingredient of Salvia miltiorrhiza Bunge, on the proliferation, metastasis and CD40-AKT-NF-κB signalling pathway in endometrial carcinoma (EC). Human EC cell lines (Ishikawa and HEC-1A) were treated with varying concentrations of SA, CD40 soluble ligand (sCD40L) or a combination of both. Cell viability, proliferation, invasion and migration were assessed using MTT, colony formation and transwell assays. Flow cytometry was used to analyse apoptosis and cell cycle progression. qRT-PCR evaluated the mRNA level of CD40. The protein expression of CD40, p-AKT, p-mTOR, p-p65, and p52 was evaluated via Western blot and immunofluorescence. A subcutaneous tumour model was used to examine the impact of SA on tumour growth, followed by immunohistochemical analysis of Ki-67, CD40, p-AKT and p-mTOR. SA treatment reduced EC cell viability, proliferation, invasion and migration, while also triggering apoptosis and inducing cell cycle arrest in the G0/G1 phase in a dose-dependent way. These effects correlated with marked downregulation of CD40, p-AKT, p-mTOR, p-p65 and p52 expression. Conversely, activation of CD40 signalling with sCD40L promoted EC cell malignancy and overturned the anti-tumour effects of SA on EC cells. Additionally, SA treatment suppressed tumour growth in xenograft mouse models, along with reduced levels of Ki67, CD40, p-AKT, p-mTOR, p-p65 and p52 in mouse tumour tissues, which were counteracted by sCD40L co-treatment. SA effectively suppresses endometrial carcinoma progression by targeting the CD40-AKT-NF-κB pathway.
我们旨在研究丹参活性成分丹酚酸A（SA）对子宫内膜癌（EC）增殖、转移及CD40-AKT-NF-κB信号通路的影响。用不同浓度的SA、CD40可溶性配体（sCD40L）或两者组合处理人EC细胞系（Ishikawa和HEC-1A）。采用MTT法、集落形成实验和Transwell实验评估细胞活力、增殖、侵袭和迁移能力。运用流式细胞术分析细胞凋亡和细胞周期进程。通过qRT-PCR检测CD40的mRNA水平。采用蛋白质免疫印迹法和免疫荧光法评估CD40、p-AKT、p-mTOR、p-p65和p52的蛋白表达。利用皮下肿瘤模型研究SA对肿瘤生长的影响，随后对Ki-67、CD40、p-AKT和p-mTOR进行免疫组化分析。SA处理可降低EC细胞活力、增殖、侵袭和迁移能力，同时以剂量依赖方式诱导细胞凋亡并使细胞周期停滞于G0/G1期。这些作用与CD40、p-AKT、p-mTOR、p-p65和p52表达的显著下调相关。相反，用sCD40L激活CD40信号可促进EC细胞恶性增殖并逆转SA对EC细胞的抗肿瘤作用。此外，SA处理可抑制异种移植小鼠模型中的肿瘤生长，同时降低小鼠肿瘤组织中Ki67、CD40、p-AKT、p-mTOR、p-p65和p52的水平，而sCD40L联合处理可抵消这些作用。SA通过靶向CD40-AKT-NF-κB通路有效抑制子宫内膜癌进展。
