Mechanisms Of ITE’s Effects On Glioma Cell Migration And Proliferation

One of the drug target of GBM immunotherapy research is the enzyme indoleamine2,3-dioxygenase/Tryptophan-2,3-dioxygenase,(IDO/TDO)metabolizing tryptophane(Try)to kynurenine(Kyn).2-(1’H-indole-3’-carbonyl)-thiazole-4-carboxylic acid methyl ester(ITE)is an endogenous ligand for aryl hydro carbon receptor(AhR),we hypothesis that ITE can compete with kynurenine for binding to AhR and block this pathway.Our unpublished data shown that ITE can inhibit the migration and invasion of GBM.We study ITE’s effects on two commonly used glioma cell lines U87MG and GL261.It was found that ITE had no significant effect on the proliferation of U87MG,and in GL261 had a tendency to promote cell proliferation.Time-lapse photography showed that U87MG cells spread less on the plastic culture suferce and detached rounded up faster,suggesting that ITE can reduce cell adhesion reduction.In addition,in 2D cell culture ITE significantly reduced the number of β-actin bunldes at the cell edge suggested that ITE significantly reduced the number stable filopodia.In 3D cell culture ITE weakened the filopodia tip to the collagen matrix.The mechanism was under investigation.Futher studies revealed that the proliferation-related genes CDK4 and PRMT5 were enriched in the protein complex of U87MG that was down-regulated by ITE,but not significantly enriched in GL261.Real-Time PCR verified the corresponding changes in CDK4 and PRMT5.It was found that the change of the two internal parameters of ACTB was consistent with the transcriptome data.The PCR results of HPRT,GAPDH and ACTB were different from the transcriptome structure.Both results partially explain the observed differences in proliferation,suggesting that ITE-AHR has different effects on human and murine glioma cell proliferation-related pathways.We also performed pathway and GO enrich hment and protein interaction/protein complex analysis of ITE-regulated genes,as well as mRNA-miRNA integration analysis.It was found that different analysis strategies provide complementary information;We found that different transcriptome analysis strategies provide complementary information.The ITE-regulated U87MG and GL261 transcriptome data have similarities and differences,suggesting that multiple models should be integrated when using cell line models for scientific research,The differentially expressed genes(DEGs)and microRNAs of U87MG and GL261 are mostly different,suggesting that the ITE regulate gene express network differed a lot in U87MG and GL261,cautions should be taken when extrapolate mouse model date to human.It provides a reference for the study of subsequent glioma models and extrapolation of clinical response.