重组6号染色体开放阅读框120蛋白通过PI3K/Akt信号通路促进人脐静脉内皮细胞的血管生成和内皮-间充质转化。

Recombinant chromosome 6 open reading frame 120 protein promotes angiogenesis and endothelial‑to‑mesenchymal transition in human umbilical vein endothelial cells via the PI3K/Akt signaling pathway.

作者信息

Lin Yingying, Wang Xin, Li Yanyan, Cui Xinyu, Zhu Na, Li Xin

机构信息

Department of Center of Integrated Traditional Chinese and Western Medicine, Peking University Ditan Teaching Hospital, Beijing 100015, P.R. China.

Department of Center of Integrated Traditional Chinese and Western Medicine, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China.

出版信息

Mol Med Rep. 2025 Sep;32(3). doi: 10.3892/mmr.2025.13596. Epub 2025 Jun 20.

DOI:10.3892/mmr.2025.13596

PMID:40539464

Abstract

The vascular endothelium plays a pivotal role in modulating various physiological processes and its dysfunction is fundamental to the development of numerous vascular and non‑vascular diseases. Chromosome 6 open reading frame 120 (C6ORF120) has been implicated in cellular processes such as apoptosis, inflammation, immunomodulation and fibrosis. However, the specific effects of C6ORF120 on endothelial cell function remain unclear. The present study aimed to explore the potential role of C6ORF120 in endothelial dysfunction and its underlying molecular mechanisms. It synthesized recombinant C6ORF120 protein (rC6ORF120) and assessed its effects on human umbilical vein endothelial cells (HUVECs) through various functional assays, including the CCK‑8 assay for proliferation, scratch assay for migration and tube formation assay for angiogenesis. Additionally, immunofluorescence (IF) and western blotting (WB) were employed to evaluate endothelial‑mesenchymal transition (EndMT). The present study also quantified the expression of key proteins within the PI3K/Akt signaling pathway to elucidate its role in mediating the effects of rC6ORF120 on HUVECs. Treatment with rC6ORF120 significantly enhanced HUVEC proliferation (200 ng/ml vs. control at 72 h, 1.14±0.01 vs. 1.05±0.02; t=8.15; P<0.001) and induced phenotypic changes. In migration and angiogenesis assays, rC6ORF120‑treated HUVECs exhibited increased wound closure (37.69±2.74% vs. 66.16±6.13%; t=7.35; P=0.002) and angiogenesis assays showed significant improvements in tube formation parameters such as total tubule length (77,199.67±4,684.88 µm vs. 96,203.00±3,354.89 µm; t=5.71; P=0.002). WB and IF analyses both indicated that rC6ORF120 promotes EndMT in HUVECs. Furthermore, rC6ORF120 treatment increased PI3K/Akt phosphorylation significantly compared with controls (p‑PI3K; 1.57±0.18 vs. 1.00±0.00; t=5.64; P=0.005). LY294002 significantly reversed these effects on EndMT and angiogenesis (P<0.05), while the effect on cell migration was less pronounced (P=0.565). Our study highlights the critical role of C6ORF120 in HUVECs, promoting proliferation, migration, angiogenesis and EndMT, which are mediated, at least in part, by the PI3K/Akt pathway.

摘要

血管内皮在调节各种生理过程中起关键作用，其功能障碍是众多血管性和非血管性疾病发生发展的根本原因。6号染色体开放阅读框120（C6ORF120）与细胞凋亡、炎症、免疫调节和纤维化等细胞过程有关。然而，C6ORF120对内皮细胞功能的具体影响仍不清楚。本研究旨在探讨C6ORF120在内皮功能障碍中的潜在作用及其潜在分子机制。它合成了重组C6ORF120蛋白（rC6ORF120），并通过各种功能试验评估其对人脐静脉内皮细胞（HUVECs）的影响，包括用于增殖的CCK-8试验、用于迁移的划痕试验和用于血管生成的管形成试验。此外，采用免疫荧光（IF）和蛋白质印迹法（WB）评估内皮-间充质转化（EndMT）。本研究还对PI3K/Akt信号通路中的关键蛋白表达进行了定量，以阐明其在介导rC6ORF120对HUVECs作用中的作用。用rC6ORF120处理显著增强了HUVECs的增殖（72小时时200 ng/ml与对照相比，1.14±0.01对1.05±0.02；t=8.15；P<0.001）并诱导了表型变化。在迁移和血管生成试验中，经rC6ORF120处理的HUVECs伤口闭合增加（37.69±2.74%对66.16±6.13%；t=7.35；P=0.002），血管生成试验显示管形成参数如总小管长度有显著改善（77199.67±4684.88 µm对96203.00±3354.89 µm；t=5.71；P=0.002）。WB和IF分析均表明rC6ORF120促进HUVECs中的EndMT。此外，与对照相比，rC6ORF120处理显著增加了PI3K/Akt磷酸化（p-PI3K；1.57±0.18对1.00±0.00；t=5.64；P=0.005）。LY294002显著逆转了这些对EndMT和血管生成的影响（P<0.05），而对细胞迁移的影响不太明显（P=0.565）。我们的研究突出了C6ORF120在HUVECs中的关键作用，促进增殖、迁移、血管生成和EndMT，这些至少部分由PI3K/Akt途径介导。