The Influence Of Interleukin 17 On The Proliferation And Migration Of Glioma Cells And The Molecular Mechanisms

Objective: Interleukin 17(IL?17),as a pro-inflammatory cytokine,is up-regulated in the sera and tumor tissues of glioma patients;however the effects of IL-17 on glioma proliferation and migration remain unclear.In this study,IL-17 role and its potential mechanisms in the proliferation and migration of glioma cells was determined.Methods:(1)The expression of IL-17 RA on SHG-44 and U373 cells was evaluated by Flow CytoMetry.Using EdU kit examine the effect of IL-17 on the proliferation of SHG-44 cells and U373 cells.The influence of IL-17 on the proliferation and migration of these two types glioma cells evaluated by colony formation assay and cell scratch assay.Evaluating the ability of IL-17 to induce tumor formation of glioma cells in vivo experiments: SHG-44 cells were cultured and incubated with or without 50 ng/ml of IL-17 for 15 min,and then injected into nude mice(n=6)subcutaneously for tumor growth in vivo until 3 weeks,tumor sizes were detected at different time points and tumor weights were obtained at last time points.(2)The antibody chips of intracellular signaling array were used to screen some signaling molecules in the IL-17-induced glioma cells.Confirming the results of antibody chips experiment and detect the other related signaling molecules by western blot.(3)SHG-44,U373 cell lines were designed into inhibition group and small interfering RNA group,namely DMSO group,50 ng/mL IL-17 treated group,IL-17-treated + LY294002 group,IL-17-treated + Perifosine group,IL-17-treated + BAY11-7082 group,and control group,IL-17-treated + siCTR group,IL-17-treated + siPI3 K group,IL-17-terated + siAkt1 group,IL-17-treated + siP65 group.Confirming the upstream and downstream relationship among this signaling molecules in the glioma cell of the two groups by western blot.SHG-44 cells of inhibition group were injected into nude mice(n=8)subcutaneously for tumor growth in vivo until 3 weeks,tumor sizes were detected at different time points and tumor weights were obtained at last time points.Results:(1)Flow cytometry data showed that both SHG-44 and U373 cell lines significantly expressed IL-17 R.The results of EdU detection and colony formation assay showed that IL-17 could markedly increase the proliferation of SHG-44 cells and U373 cells.It was found that IL-17 could improve the migration of not only SHG-44 cells but also U373 cells by cell scratch assay.The results of in vivo experiment revealed that IL-17 incubation could obviously enhance the tumor formation of glioma cells in nude mice.(2)The results of antibody chips demonstrated that IL-17 could markedly increase the level of p-Akt1 in glioma cells.It also found that IL-17 stimulation obviously increased the phosphorylation of NF-?B-p65 in glioma cells.(3)PI3K/Akt1 is located upstream of NF-?B-p65.The results of EdU detection,colony formation assay and scratch assay indicate that the activation of PI3K/Akt1/NF-?B-p65 axis contributes to the proliferation and migration of glioma cells exposed to IL-17.In vivo experiment,the data showed that IL-17 could induce the tumor formation of glioma cells through the activation of PI3K/Akt1/NF-?B-p65 axis.Conclusion:(1)IL-17 can induces the proliferation and migration of glioma cells.(2)IL-17 promotes the proliferation and migration of glioma cells via PI3K/Akt1/NF-?B-p65 activation.