摘要目的:探讨细胞周期相关肿瘤高表达蛋白（CREPT）在胶质瘤血管内皮细胞中的表达特征及其调控血管生成的作用机制。方法:基于人胶质瘤样本单细胞测序数据，分析CREPT在不同细胞类型中的表达模式；采用免疫组织化学与免疫荧光染色验证其在胶质瘤组织中的表达分布；分析CREPT表达与胶质母细胞瘤微血管密度的相关性；通过基因集富集分析评估CREPT对内皮细胞功能的影响；构建CREPT敲低的人脑微血管内皮细胞模型，并通过成管实验评估CREPT对血管形成能力的影响；利用Gilovis数据库分析CREPT与血管内皮生长因子A的相关性、分析两者在胶质母细胞瘤不同区域的表达模式，并通过免疫组织化学进行验证；最后，通过Western blot检测黏附连接蛋白及下游信号通路分子的表达变化。结果:CREPT在胶质瘤血管内皮细胞中特异表达，高表达CREPT组的微血管密度显著高于低表达CREPT组（ t=2.083， P<0.05）；基因集富集分析结果显示CREPT参与调控内皮细胞成管与黏附连接功能，敲低CREPT可显著抑制人永生化脑微血管内皮细胞（hCMEC）的成管能力（ F=12.99， P<0.05）；差异表达分析与临床样本验证显示，CREPT与血管内皮生长因子A在假栅栏样坏死等区域表达均升高（ t=7.513， P<0.05； t=9.461， P<0.001），且两者呈显著正相关（ r=0.36， P<0.05）；敲低CREPT后，血管内皮生长因子受体2表达显著下降（ F=12.74， P<0.05）。 结论:CREPT在胶质瘤血管内皮细胞特异表达，通过调控血管内皮生长因子A-血管内皮生长因子受体2信号通路，增强胶质瘤血管内皮细胞的成管能力，进而促进肿瘤血管生成，为胶质母细胞瘤抗血管治疗提供了新的潜在靶点。

abstractsObjective:To investigate the expression pattern of cell cycle-related and expression-elevated protein in tumor (CREPT) in vascular endothelial cells of glioma and to elucidate its mechanism in regulating angiogenesis.Methods:Based on single-cell sequencing data from human glioma samples, CREPT expression patterns across different cell types were analyzed. Immunohistochemistry and immunofluorescence staining were used to examine its expression and distribution in glioma tissues. The correlation between CREPT expression and micro-vessel density (MVD) in glioblastoma was evaluated. Gene set enrichment analysis (GSEA) was performed to assess CREPT′s impact on endothelial cell functions. A CREPT-knockdown hCMEC model was established. Tube formation assays were conducted to evaluate CREPT′s effect on angiogenic capacity. The Gilovis database was used to analyze the correlation between CREPT and vascular endothelial growth factor A (VEGFA), and to validate their expression patterns in different glioblastoma regions, which was further confirmed using immunohistochemistry. Western blot was also performed to detect changes in the expression of adhesion junction proteins and downstream signaling molecules.Results:CREPT was specifically expressed in glioma vascular endothelial cells, and the micro-vessel density (MVD) in the group with high CREPT expression was significantly higher than that in the group with low CREPT expression ( t=2.083, P<0.05). GSEA showed that CREPT was involved in regulating endothelial cell tube formation and adhesion junction functions. The tube-forming ability of hCMEC was significantly inhibited by knockdown of CREPT ( F=12.99, P<0.05). Using differential expression analysis and clinical sample validation, it was revealed that both CREPT and VEGFA were upregulated in regions such as pseudopalisading necrosis ( t=7.513, P<0.05; t=9.461, P<0.001), and their expression levels were positively correlated ( r=0.36, P<0.05). After CREPT knockdown, the expression of VEGFR2 was markedly decreased ( F=12.74, P<0.05). Conclusions:CREPT is specifically expressed in glioma vascular endothelial cells. It also enhances the tube-forming capacity of glioma vascular endothelial cells and promotes tumor angiogenesis by regulation of the VEGFA-VEGFR2 signaling pathway. These findings provide a novel potential target for anti-angiogenic therapy in glioblastoma.