ABSTRACT
OBJECTIVE@#To investigate the effect of interference of P2X4 receptor expression in tumor-associated macrophages (TAMs) on invasion and migration of glioma cells.@*METHODS@#C57BL/6 mouse models bearing gliomas in the caudate nucleus were examined for glioma pathology with HE staining and expressions of Iba-1 and P2X4 receptor with immunofluorescence assay. RAW264.7 cells were induced into TAMs using conditioned medium from GL261 cells, and the changes in mRNA expressions of macrophage polarization-related markers and the mRNA and protein expressions of P2X4 receptor were detected with RT-qPCR and Western blotting. The effect of siRNA-mediated P2X4 interference on IL-1β and IL-18 mRNA and protein expressions in the TAMs was detected with RT-qPCR and Western blotting. GL261 cells were cultured in the conditioned medium from the transfected TAMs, and the invasion and migration abilities of the cells were assessed with Transwell invasion and migration experiment.@*RESULTS@#The glioma tissues from the tumor-bearing mice showed a significantly greater number of Iba-1-positive cells, where an obviously increased P2X4 receptor expression was detected (P=0.001), than the brain tissues of the control mice (P < 0.001). The M2 macrophage markers (Arg-1 and IL-10) and M1 macrophage markers (iNOS and TNF-α) were both significantly up-regulated in the TAMs derived from RAW264.7 cells (all P < 0.01), but the up-regulation of the M2 macrophage markers was more prominent; the expression levels of P2X4 receptor protein and mRNA were both increased in the TAMs (P < 0.05). Interference of P2X4 receptor expression significantly lowered the mRNA(P < 0.01)and protein (P < 0.01, P < 0.05)expression levels of IL-1β and IL-18 in the TAMs and obviously inhibited the ability of the TAMs to promote invasion and migration of the glioma cells (P < 0.05).@*CONCLUSION@#Interference of P2X4 receptor in the TAMs suppresses the migration and invasion of glioma cells possibly by lowering the expressions of IL-1β and IL-18.
Subject(s)
Animals , Mice , Culture Media, Conditioned , Glioma , Interleukin-18 , Mice, Inbred C57BL , RNA, Messenger , Receptors, Purinergic P2X4/metabolism , Tumor-Associated MacrophagesABSTRACT
Objective To provide evidence for the relationship between TMD and occlusal trauma by studying the mechanics influence and displacement of condyle by building the three dimensional finite element analysis model andstimulating the abnormal bite force leading occlusal trauma.Methods We collected DICOM date of healthy male volunteers with normal occlusal relationship using CBCT scan with constraint on condylar top and formed the finite model with divide mesh.Two conditions were built: (1) We operated different vertical load on the left mandibular molar occlusal surface; (2) We operated different buccal direction load on the same modelto study the mechanic change on left condylar.Results When the three dimensional finite model of mandible was built, the mechanic region of condyle stayed the same but the stress and tension increased accordingly as the loda duplicate rose. As the same buccal operation load was operated,the stress and tension became larger and the displacement became longer.Conclusion The abnormal bite force causing occlusal trauma can change mechanic conduct on condyle and lead to the pathological change of condyle.