From the Departments of Dermatology, Plastic and Reconstructive Surgery, and Laser and Aesthetic Medicine, Shanghai Ninth People's Hospital, and the Shanghai Institute of Immunology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine.
Plast Reconstr Surg. 2021 Mar 1;147(3):412e-423e. doi: 10.1097/PRS.0000000000007672.
Hypoxia may play a role in the pathogenesis of infantile hemangioma. Cysteine-rich angiogenic inducer 61 (Cyr61), or CCN1, can be induced under hypoxic conditions in several types of cells. However, whether CCN1 has any impact on infantile hemangioma remains unknown. This study aims to explore the expression of CCN1 in infantile hemangioma and to investigate the effect of hypoxia on CCN1 and vascular endothelial growth factor-A (VEGF-A) production.
Hemangioma-derived endothelial cells and hemangioma-derived stem cells were isolated from surgical specimens of proliferative infantile hemangioma. RNA extracted from infantile hemangioma tissue, hemangioma-derived endothelial cells, and hemangioma-derived stem cells was used to analyze gene expression by real-time polymerase chain reaction. The effects of CCN1 blockade were examined in hemangioma-derived stem cells. Immunostaining, immunoblotting, and enzyme-linked immunosorbent assays were used to assess protein expression.
By double-label immunofluorescence staining, the authors first identified that CCN1 was abundant in proliferative infantile hemangioma lesions and colocalized well with immature microvessels. The authors found that the mRNA level of CCN1 in proliferative infantile hemangioma was significantly higher than in healthy controls, as was involuting infantile hemangioma. Treatment with the hypoxia inducer cobalt chloride dramatically increased CCN1 production in hemangioma-derived endothelial cells in a time-dependent manner. Furthermore, blocking or knockdown of CCN1 expression reduced the expression of VEGF-A in hemangioma-derived stem cells. Lastly, the signaling pathway study showed that CCN1 up-regulation of VEGF-A synthesis in hemangioma-derived stem cells depends on nuclear factor-κB and JNK activation.
These findings provide new evidence that CCN1 participates in the crosstalk between hemangioma-derived endothelial cells and hemangioma-derived stem cells through promoting VEGF-A expression in the hypoxic environment of infantile hemangioma angiogenesis and vasculogenesis. Targeting of CCN1 might be a novel therapeutic strategy for infantile hemangioma.
缺氧可能在婴儿血管瘤的发病机制中起作用。富含半胱氨酸的血管生成诱导因子 61（Cyr61）或 CCN1 可在几种类型的细胞中在缺氧条件下诱导。然而，CCN1 是否对婴儿血管瘤有任何影响尚不清楚。本研究旨在探讨 CCN1 在婴儿血管瘤中的表达，并研究缺氧对 CCN1 和血管内皮生长因子-A（VEGF-A）产生的影响。
从增殖期婴儿血管瘤的手术标本中分离出血管内皮细胞和血管瘤衍生的干细胞。从婴儿血管瘤组织、血管瘤衍生的内皮细胞和血管瘤衍生的干细胞中提取 RNA，通过实时聚合酶链反应分析基因表达。在血管瘤衍生的干细胞中检查 CCN1 阻断的效果。免疫染色、免疫印迹和酶联免疫吸附试验用于评估蛋白质表达。
通过双标记免疫荧光染色，作者首次鉴定出 CCN1 在增殖期婴儿血管瘤病变中含量丰富，与不成熟的微血管很好地共定位。作者发现，增殖期婴儿血管瘤中 CCN1 的 mRNA 水平明显高于健康对照组和消退期婴儿血管瘤。缺氧诱导剂氯化钴处理可显著增加血管瘤衍生内皮细胞中 CCN1 的产生，呈时间依赖性。此外，阻断或敲低 CCN1 表达可降低血管瘤衍生干细胞中 VEGF-A 的表达。最后，信号通路研究表明，CCN1 通过激活核因子-κB 和 JNK 上调血管瘤衍生干细胞中 VEGF-A 的合成。
这些发现为 CCN1 通过在婴儿血管瘤血管生成和血管发生的缺氧环境中促进 VEGF-A 表达参与血管瘤衍生内皮细胞和血管瘤衍生干细胞之间的串扰提供了新的证据。针对 CCN1 可能是治疗婴儿血管瘤的一种新的治疗策略。
