PXN and TRIO are elevated in CD133+GSCs transcriptomically
Authors: Sunguroglu Asuman , Bunsuz Merve , Yukselten Yunus , Bal Merve Gulsen , Ozkanca Seyma , Yildiz Dilara Akcora , Demircan Kadir , Ugur Hasan Caglar 
Published: November 2023

OBJECTİVEGlioblastoma Multiforme (GBM) is one of the highly aggressive and malign tumor and associated with cancer related deaths of worldwide. Glioblastoma stem cells (GSCs) are playing essential role in tumor progression and invasiveness in brain. GSCs migrate non‐cancerous tissues and promote cancer expansion. According to cancer stem cell hypothesis, it is assumed that cancer stem cells are responsible for tumor initiation, metastasis, and resistance to treatments. CD133, also known as prominin‐1 is a transmembrane glycoprotein, is identified as a GSCs surface marker in GBM. The aim of this research was that to identify cell adhesion molecules, which are expressed particularly at GSCs and investigate therapeutic targets to eradicate GSCs in GBM.Integrin signalling pathway and downstream adaptors are considerable in cell migration and invasion of cancer cells to non‐cancerous tissues. Elevated expression of integrin adaptors are shown in many cancers, also in GBM. PXN is a focal adhesion‐associated protein and serves as an adaptor protein to recruit downstream signaling molecules. Paxillin is associated with focal adhesion destabilization via microtubule‐binding and promotes cell motility and spreading. TRIO is serves as Rho guanine nucleotide exchange factor in cell and promotes cell migration through actin re‐organization. RAC proteins as a small GTPase regulate various cellular events, including the cell growth and cytoskeletal reorganization. Rac‐activating guanine nucleotide exchange factor TRIO, directly associated with RAC activation and cytoskeletal reorganization.MATERIAL METHODSGBM primary cells were freshly isolated from human glioblastoma tissue samples and cultured in DMEM High glucose supplemented with 10% fetal calf serum and 1% penicillin‐streptomycin at 37 °C in 5% CO2‐humidified incubator. We have isolated CD133+ cells from GBM primary cells obtained from 10 GBM patients by MACS method using AC133 CD133 antibody. Following RNA isolation from CD133 + and CD133− cells, cDNA synthesis was performed. Finally according to micro array protocol, cell adhesion array was applied. Gene expression levels were analyzed using the ‐delta delta Ct method. One‐sample t‐test was used to determine the mean fold changes (increase/decrease) of genes. Statistical analysis was performed using SPSS software for windows version 13.0.RESULTSAs a result of this study, PXN (Paxillin) and TRIO (Triple functional domain (PTPRF interacting)) genes are found upregulated in CD133+ GCSs compared CD133− cells 3.18 and 4.25 fold, respectively (p<0.05).CONCLUSIONIn summary, we found that PXN and TRIO genes have elevated expression levels specifically in CD133+ cells and both of them are related with cell migration and invasion in cellular process. In the light of these results PXN and TRIO might be a useful marker to target GSCs and repression of these genes might diminish cellular invasion of GBM cells. Further studies are required to verifying therapeutic effect of these target molecules in GBM treatment.This research has been supported by The Scientific and Technological Research Council of Turkey (No:114S189).