Reactions Of Microglial Polarization On Glioma Cells

Objective:Human glioma cells U251 and U87 were cultured in different conditioned medium after human microglia HMC-3 was induced by different inducers.The cell migration,cell invasion,cell proliferation and morphological changes of cultured human glioma cells U251 and U87 were observed to provide evidence for exploring the action mechanism of microglia in glioma microorganisms.Methods:1.Cultured human microglia cells HMC-3,human glioma cells U251 and human glioma cells U87 Induction of human microglia HMC-3 into microglia HMC-3 activated state with LPS(M1)Induction of human microglia HMC-3 into microglia HMC-3activated state with IL-4(M2)and collection of different groups of CM: control group(NC group)lipopolysaccharide induced group(LPS group)and interleukin-4 induced group(IL-4 group)The morphological changes of human microglia HMC-3 in resting state(M0),microglia HMC-3 in activated state(M1)and microglia HMC-3 in activated state(M2)were observed under inverted optical microscope Human glioma cell line U251 and human glioma cell line U87 were cultured with different conditioned medium of human microglia(HMC-3-CM).The morphology of human glioma cell line U251 and human glioma cell line U87 was observed under inverted optical microscope.2.Two subtypes of human microglia HMC-3 induced from resting M0 to M1 M2 were identified by flow cytometry.The secretion of IL-6 and TNF-α in LPS group was detected by ELISA.The secretion of IL-10 and TGF-β in IL-4 group was also detected by ELISA.3.Human microglia cells were co-cultured with human glioma cell U251 and human glioma cell U87 in different conditioned media,and then the effects of HMC-3-CM on the migration,invasion and proliferation of human glioma cell U251 and human glioma cell U87 were detected by cell scratch test,Transwell migration and invasion test and MTT test,and the results were statistically analyzed.Results:1.The morphology of M1 M2 subtype cells produced by human microglia HMC-3induced by LPS IL-4 changed from round or oval resting state to rod-shaped active state with enlarged cell body;The morphology of some activated microglia was amoeba-like Human glioma cell line U251 U87 was reduced in size by different conditioned medium produced by human microglia HMC-3 induced by LPSIL-4,and its morphology changed from long rod shape to round or oval shape.2.The distribution percentage of M1-type cells after HMC-3 polarization induced by LPS increased significantly and the distribution percentage of M2-type cells after HMC-3 polarization induced by IL-4 increased significantly.The secretion of IL-6 TNF-αand other M1 cytokines in human microglia induced by LPS were significantly increased and there was statistical significance.The secretion of IL-10 TGF-β and other M2 cytokines in human microglia induced by LPS were not statistically significant;The expression of IL-10 TGF-β and other M2 cytokines in HMC-3 cells induced by IL-4 was significantly higher than that in HMC-3 cells induced by IL-4,and the expression of IL-6TNF-α and other M1 cytokines in HMC-3 cells was significantly higher than that in HMC-3 cells induced by IL-4.3.The cell mobility of human glioma cell U251 U87 in LPS group was statistically significant and the cell mobility of human glioma cell U251 U87 in IL-4 group was significantly increased;Human glioma cells U251 U87′s migration quantity in LPS group was statistically significant and human glioma cells U251 U87′s migration quantity in IL-4 group was significantly increased;The absorbance(490nm)of human glioma cell U251 U87 in LPS group was statistically significant;The absorbance(490nm)of human glioma cells U251 U87 in IL-4 group was statistically significant.Conclusion:1.LPS induced the polarization of HMC-3 mainly to human microglia M1 type,and IL-4 induced the polarization of HMC-3 mainly to human microglia M2 type.2.LPS induced human microglia HMC-3 to mainly secrete M1-type cytokines such as IL-6 and TNF-α,and IL-4 induced HMC-3 to mainly secrete M2-type cytokines such as IL-10 and TGF-β.3.LPS induced CM of HMC-3 inhibited human glioma cells U251 U87′s migration,invasion and proliferation.IL-4 induced CM of HMC-3 can promote human glioma cells U251 U87′s migration,invasion and proliferation.