He Yang, Lin Jian, Li Yi, Cheng Xiaobo, Wang Tong, Wang Wei, Zeng Weixing, Li Yongsheng
Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China; The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650031, China; Shiyan Key Laboratory of Comprehensive Prevention and Treatment of Oral Cancer, Department of Stomatology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China; Institute of Oral Diseases, School of Dentistry, Hubei University of Medicine, Shiyan, Hubei 442000, China.
Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China; The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650031, China; Kunming Stomatological Hospital North City District Branch, 650231, China.
Microvasc Res. 2025 Sep;161:104833. doi: 10.1016/j.mvr.2025.104833. Epub 2025 Jun 24.
This study aims to investigate the regulatory role and underlying molecular mechanisms of DNA Damage Inducible Transcript 4 (DDIT4) in the pathogenesis of Venous Malformations (VMs), providing foundational experimental evidence for potential targeted therapies.
Bioinformatic analysis identified DDIT4 as a key differentially expressed gene in VMs, and the sgGSEA method was employed to predict its potential biological functions. Immunohistochemical staining and immunofluorescence were performed to validate the expression level of DDIT4 and its association with vascular density. A lentiviral VMs cell model was established to assess DDIT4 expression levels. The effects of DDIT4 knockdown on VMs cell function were evaluated, with mechanistic insights gained through transcriptome sequencing and Western blot analysis. Further validation was performed using 3D VMs cell models and nude mouse xenografts with DDIT4 knockdown. Additionally, exogenous functional rescue experiments were conducted by activating the NF-κB pathway with lipopolysaccharide (LPS) in DDIT4 knockdown VMs 3D cell models and nude mouse xenografts to further investigate the role of DDIT4.
DDIT4 was upregulated in VMs tissues and correlated with angiogenesis. DDIT4 knockdown increased cell roundness, inhibited proliferation, migration, and NF-κB pathway activation, and blocked angiogenesis in VMs 3D models and lesion formation in nude mouse xenografts, while suppressing the NF-κB pathway in both. NF-κB pathway activation restored angiogenesis in both models.
DDIT4 knockdown inhibits VMs progression by suppressing the NF-κB pathway, suggesting that DDIT4 may serve as a potential therapeutic target.
本研究旨在探讨DNA损伤诱导转录本4（DDIT4）在静脉畸形（VMs）发病机制中的调控作用及潜在分子机制，为潜在的靶向治疗提供基础实验依据。
生物信息学分析确定DDIT4为VMs中关键的差异表达基因，并采用单样本基因集富集分析（sgGSEA）方法预测其潜在生物学功能。进行免疫组织化学染色和免疫荧光以验证DDIT4的表达水平及其与血管密度的关系。建立慢病毒VMs细胞模型以评估DDIT4表达水平。评估DDIT4敲低对VMs细胞功能的影响，并通过转录组测序和蛋白质免疫印迹分析获得作用机制方面的见解。使用3D VMs细胞模型和DDIT4敲低的裸鼠异种移植模型进行进一步验证。此外，通过在DDIT4敲低的VMs 3D细胞模型和裸鼠异种移植模型中用脂多糖（LPS）激活NF-κB通路进行外源性功能挽救实验，以进一步研究DDIT4的作用。
DDIT4在VMs组织中上调并与血管生成相关。DDIT4敲低增加细胞圆度，抑制增殖、迁移和NF-κB通路激活，并阻断VMs 3D模型中的血管生成和裸鼠异种移植模型中的病变形成，同时在两者中均抑制NF-κB通路。NF-κB通路激活恢复了两种模型中的血管生成。
DDIT4敲低通过抑制NF-κB通路抑制VMs进展，提示DDIT4可能作为潜在的治疗靶点。
