The Regulation Pathway of VEGF Gene Expression Is Different between 2D Cells and 3D Spheroids in Human Lung Cancer Cells

Abstract

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. Hypoxiainducible 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.

Therapeutic effects and underlying mechanism of poly (L-glutamic acid)-g-methoxy poly (ethylene glycol)/combretastatin A4/BLZ945 nanoparticles on Renca renal carcinoma

“…In essence, the increase in HIF-1α appears to be associated with the decrease in VEGFA expression. This conclusion aligns with the results reported by Ryo Onodera (Onodera et al, 2023).…”

Potential therapeutic target secretogranin II might cooperate with hypoxia‐inducible factor 1α in sunitinib‐resistant renal cell carcinoma

“… 23 Additionally, it has been reported that the VEGF gene expression pathways differ between 2D cell cultures and 3D spheroids, and in certain scenarios, increased HIF1α levels are associated with reduced VEGFA expression. 24 These factors could potentially explain the lack of significant results in the tube formation assay following SCG2 overexpression (Figure 6E ). Considering the intricate nature of disease processes and the involvement of various factors, if there is indeed an association between elevated HIF1α and decreased VEGFA, it is probable that additional regulatory mechanisms and modulators contribute to this relationship.…”

Three‐dimensional spheroid formation of adipose‐derived stem cells improves the survival of fat transplantation by enhance their therapeutic effect

“…[10,11] Studies have shown that the hypoxic environment of the cells in the middle of the spheroids can not only enhance their adaptation to ischemic tissue, [12] but also promote the secretion of more cytokines, such as VEGF, bFGF, and FGF2. [13] In a nude mouse model, scholars have also found that ADSCs spheroids have a higher retention rate than monolayer cells and have a better potential for treatment by intramuscular injection of ADSCs. [14] In our study, we tested whether 3D spheroid culture could regulate autophagy, apoptosis, senescence, and secretion of cytokines and further analyzed the influence of spheroid formation on neovascularization and the survival of transplanted fat particles.…”

“…[ 10,11 ] Studies have shown that the hypoxic environment of the cells in the middle of the spheroids can not only enhance their adaptation to ischemic tissue, [ 12 ] but also promote the secretion of more cytokines, such as VEGF, bFGF, and FGF2. [ 13 ] In a nude mouse model, scholars have also found that ADSCs spheroids have a higher retention rate than monolayer cells and have a better potential for treatment by intramuscular injection of ADSCs. [ 14 ]…”