• Adenocarcinoma
• epithelial growth factor (EGFR)
• 3D model

Adenocarcinoma is the most common isotype of lung cancer and presents EGFR mutations in 17% of cases. Patients with lung cancer that harbor EGFR mutations benefit from specific tyrosine kinase inhibitors treatment. The combination of EGFR tyrosine kinase inhibitors with anti-angiogenic drugs is a promising research field to enhance patients survival. Therefore, we attempt to develop an 3D in-vitro model to evaluate angiogenesis induced by EGFR mutant lung cancer cell lines and evaluate a process known as vasculogenic mimicry. We attempted to form spheroids from EGFR mutated lung cancer cells and to form tube-like structure on Geltrex matrix. Only H1975 cells were able to grow in spheroids but they did not sprout into tube-like structures while all cells lines formed structures that could resemble vascular features. Finally, we grew human vascular endothelial cells with conditioned media harvest from EGFR mutated lung cancer cell lines cultured both under standard and hypoxic conditions. In hypoxic conditions, human vascular endothelial cells grew in tube-like structures while in standard conditions grew only if cultured with PC9 and H1975 but not with HCC827 and H1650 media. These results suggested that PC9 and H1975 stimulate constitutively angiogenesis in vitro through the secretion of a soluble mediator. We compared also PC9 and H1975 versus HCC827 and H1650 gene expression using publicly available RNA sequencing data. In PC9 and H1975 cells, CDH13 AGGF1 and NOTCH4 mRNA were up regulated whereas ANGPTL3, COL4A2, EMCN and EPGN transcripts were down regulated. The over expression of AGGF1 is a promising candidate to induce tube-like formation of human vascular endothelial cells. We will validate this observation in our cell lines in further experiments.