Interference of P2X4 receptor expression in tumor-associated macrophages suppresses migration and invasion of glioma cells.
10.12122/j.issn.1673-4254.2022.05.05
- Author:
Xue Zhi YANG
1
;
Hong SHEN
2
;
Qun LI
3
;
Zi Chao DAI
4
;
Rong Qiang YANG
4
;
Guo Bin HUANG
4
;
Rui CHEN
4
;
Fang WANG
1
;
Jing Ling SONG
5
;
Hai Rong HUA
1
Author Information
1. Department of Pathology and Pathophysiology, Kunming Medical University, Kunming 650500, China.
2. Department of Pathology, Zhaotong First People's Hospital, Zhaotong 657099, China.
3. Clinic Skill Center, Kunming Medical University, Kunming 650500, China.
4. Institute of Stomatology, Affiliated Stomatology Hospital, Kunming Medical University, Kunming 650500, China.
5. Electron Microscope, Kunming Medical University, Kunming 650500, China.
- Publication Type:Journal Article
- Keywords:
P2X4 receptor;
glioma;
invasion;
migration;
tumor associated macrophages
- MeSH:
Animals;
Culture Media, Conditioned;
Glioma;
Interleukin-18;
Mice;
Mice, Inbred C57BL;
RNA, Messenger;
Receptors, Purinergic P2X4/metabolism*;
Tumor-Associated Macrophages
- From:
Journal of Southern Medical University
2022;42(5):658-664
- CountryChina
- Language:Chinese
-
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.
