The non-neuronal cholinergic system induces the inhibition in the vegf production in a glioblastoma multiforme u251 cell line

INFANTE, A; SAIBENE, P; ROSSO, D; ROSSATO, P; CANDOLFI, M; ITURRIZAGA, J; ABELLEYRO, M; JANCIC, C; VERMEULEN, M; VILLAVERDE M. S.; SALAMONE, G

Congreso; SAIC 2022; 2022

Glioblastoma multiforme (GBM) is the most common and deadly cancer of the central nervous system. This is at least in part due to the complex interaction of GBM cells with the brain microenvironment and their tendency to infiltrate normal brain tissue. This tumor is characterized by the increase in vascularity due to hypoxic regions of tumor driven by VEGF that stimulate the angiogenic activity and tumorigenesis. Acetylcholine (ACh) is a neurotransmitter, which can also modulate cell survival, proliferation and differentiation in neuronal and non-neuronal cells such as immune cells and different tumors. The aim of this work was to evaluate the influence of ACh in the growing of tumor. We examined the expression and function of ACh receptors in GBM datasets using RNA seq from Illumina HiSeq 2000 sequencing platform, by analysing GBM samples from The Cancer Genome Atlas (TCGA) using the TIMER2.0 web server. There is a decrease in the survival in the GBM patients who had increased expression of M3 cholinergic muscarinic receptors (CHRM3); we confirmed in human biopsies of tumors of patients with GBM the expression of CHRM3, by Immunohistochemistry. On the other hand, we evaluated the proliferation of U251 cell line in a 3D culture in presence of ACh for 9 days using the acid phosphatase assay (APH) and we did not find significative difference respect to the control. In addition, we evaluated the VEGF production in the supernatant of 2D (18 h) and 3D (9 d) of U251 cell cultures incubated in presence or absence of ACh by ELISA assay. Interestingly, we observed a decreased in the VEGF production in the cells cultivated with cholinergic agonist (p