血管生成在癌症的恶性转化中扮演重要角色。血管内皮生长因子（VEGF）对诱导血管生成起着重要作用。培养细胞在分析VEGF表达调控中有着重要作用，已证明VEGF表达在缺氧下被诱导。然而，在二维（2D）细胞和体内细胞之间的基因表达途径存在差异。为了解决这一问题，构建三维（3D）球状体，在3D培养中的基因表达模式更类似于体内细胞，比2D细胞更有用。本研究分析了两种人肺癌细胞株A549和H1703的3D球状体中VEGF基因表达途径。缺氧诱导因子-1α（HIF-1α）和芳香族碳氢化合物核移植物（ARNT）在3D球状体中调节VEGF基因表达。然而，在2D细胞中，VEGF基因表达并不受HIF-1α的调节。总之，我们发现，在人肺癌细胞中，2D细胞和3D球状体的VEGF基因表达调控途径不同。这些结果表明，在体内可能存在一种新的VEGF基因表达调控途径。此外，它们为分析血管生成诱导机制提供了有用的知识，也证明了3D球状体的有用性。

Angiogenesis is involved in the malignant transformation of cancers. Vascular endothelial growth factor (VEGF) is important in inducing angiogenesis. Cultured cells play an important role in analyzing the regulation of VEGF expression, and it is revealed that VEGF expression is induced under hypoxia. However, it has been shown that there are differences in the pathway for gene expression between two-dimensional (2D) cells and in vivo cells. Three-dimensional (3D) spheroids constructed in 3D culture with a gene expression pattern more similar to that of in vivo cells than 2D cells have been used to solve this problem. This study analyzed the VEGF gene expression pathway in 3D spheroids of human lung cancer cells, A549 and H1703. Hypoxia-inducible factor-1α (HIF-1α) and aryl hydrocarbon receptor nuclear translocator (ARNT) regulated VEGF gene expression in 3D spheroids. However, VEGF gene expression was not regulated by HIF-1α in 2D cells. To conclude, we found that the regulatory pathway of VEGF gene expression is different between 2D cells and 3D spheroids in human lung cancer cells. These results suggest the possibility of a new VEGF gene expression regulation pathway in vivo. In addition, they show useful knowledge for the analysis of angiogenesis induction mechanisms and also demonstrate the usefulness of 3D spheroids.