The Function And Mechnism Of Sternness Factors In CCR9/CCL25 Axis Mediated Metastasis Of T-ALL

Objective:To explore whether some sternness factors involved in the CCR9/CCL25 axis mediated metastasis of MOLT4 cells and their potential roles in metastasis of T-ALL,so as to provide scientific evidence and potential drug targets for the clinical chemotherapy as well as metastasis inhibition of T-ALL.Methods:GEO datasets of T-ALL were collected to compare the expression of CCR9 in T-ALL patients with normal control or pre-B-ALL patients,and correlation of CCR9 expression with clinical prognosis was determined by cumulative suruival analysis.GSEA was conducted to reveal the top 20 hallmarks and western blot was used to detect the PI3K/Akt/?-catenin pathway when treated with or without CCL25.Western blot or flow cytometry were used to detect the expression change of C-myc,?-catenin,Klf4,Oct4,Bmi-1,Nanog,CD34,CD38,CD44 and CXCR4 when CCR9/CCL25 axis was activated.Correlation analysis was used to detect the pertinence of CCR9 with CTNNB1,MYC,KLF4 and NANOG while cumulative suruival analysis be performed to assess the relevance of CTNNB1,MYC,KLF4 and NANOG expression with clinical recurrence.Proliferation capacity of MOLT4 cells were identified by cell counting and CCK8 assay.Impact of IWR-1-endo and 10058-F4 on MOLT4 cells was measured using MTT assay.To assess the migration and invasion of MOLT4 cells,we carried out Transwell migration and matrigel invasion assay.Results:T-ALL has a high expression of CCR9 compared with normal control or pre-B-ALL and CCR9 high expression indicates an early relapse.The PI3K/Akt/?-catenin pathway involve in the CCR9/CCL25 axis.Nuclear transcription associated sternness factors C-myc,(3-catenin and Klf4 were upregulated and Nanog was downregulated when CCR9/CCL25 axis was activated.Postive correlation between CCR9 and MYC was found,elevated expression of KLF4 promoted the survival time of T-ALL patients.Some membrane associated sternness factors,CD34,CD38,CD44 and CXCR4,showed no significant change in the CCR9/CCL25 axis.No distinct effect on the proliferation of MOLT4 cells was observed when added with CCL25 for 1 day to 5 days.B-catenin inhibitor IWR-l-endo and C-myc inhibitor 10058-F4 exhibited a potential of inhibiting proliferation of MOLT4 cells as well as a potential to inhibit migration and invasion of MOLT4 cells mediated by CCR9/CCL25 axis.Conclusion:Stemness factors C-myc,?-catenin,Klf4 and Nanog involved in the CCR9/CCL25 axis mediated metastasis of MOLT4 cells,C-myc and ?-catenin might be potential drug targets to suppress the metastasis of CCR9 high-expressing T-ALL cells.