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Objective: To investigate the effects and mechanisms of exogenous calcium-binding protein S100A4 on proliferation, survival and migration functions of glioma cells. Methods: Pan-cancer dataset was downloaded from UCSC database for the analysis of S100A4 expression and prognosis in pan-cancer, and transcriptome and clinical data of glioma patients were downloaded from CGGA database. Prognosis and progression of S100A4 expression in glioma patients were analyzed by R software. S100A4 protein network interaction of glioma patients were addressed by online analytical tools GEPIA and STRING. Human glioma cell lines (U87 and U251) were cultured in vitro, and the experiment was divided into 3 groups: PBS group, S100A4 group and S100A4+TAK242 group [Resatorvid (TAK242) was a specific inhibitor of Toll-like receptors 4 (TLR4)]. Flow cytometry was used to detect proliferation and apoptosis of glioma cells. Wound healing assay, Transwell assay and clone formation assay were used to detect migration and proliferation ability of U87 and U251 cells, and Western blot was used to detect levels of TLR4 protein and NF-κB related signaling proteins. Results: Bioinformatics results showed that S100A4 was significantly upregulated in multiple tumours (P<0.05), which included glioblastoma (GBM), lower grade glioma (LGG), stomach adenocarcinoma (STAD), liver hepatocellular carcinoma (LIHC), etc, with poorer prognosis in GBM and LGG. Compared with glioma patients with low expression of S1004, high expression S100A4 in glioma patients had shorter survival and higher degree of WHO classification. In addition, S100A4 protein in glioma patients may interact with Annexin A2 (ANXA2), TLR4 and advanced glycosylation end product-specific receptor (AGER) proteins. In cellular experiments, U87 and U251 cells showed enhanced proliferation and migration, and significantly up-regulated levels of TLR4, p-ERK1/2, p-p38 and p-p65 proteins after exogenous S100A4 treatment compared to PBS group (P<0.05), while glioma cells in S100A4+TAK242 group showed weaker proliferation and migration, and lower TLR4, p-p38 and p-p65 protein levels than those in S100A4 group, TLR4, p-ERK1/2, p-p38, p-p65 protein levels were significantly down-regulated (P<0.05). Conclusion: S100A4 may regulate function of glioma proliferation, migration and survival through TLR4/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Objective The variations in T cell subset composition and levels of inflammatory cytokines and acetylcholine receptor (AChR) antibody during the interval for myasthenia gravis (MG) patients were evaluated to investigate regulatory roles of in the pathogenesis of MG. Methods Thirty patients with MG and 20 healthy controls (HCs) were recruited. Flow cytometry was employed to detect the frequencies of CD4 + T cells, CD8 + T cells, central memory T cells (CD4 + CD45RO + CCR7 + , Tcm), effector memory T cells (CD4 + CD45RO + CCR7 - , Tem), follicular helper T cells (CD4 + CXCR5 + , Tfh) and follicular regulatory T cells (CD4 + CXCR5 + FOXP3 + , Tfr) in the peripheral blood. The levels of interleukin 2(IL-2), IL-4, IL-12, IL-17 and interferon γ (IFN-γ) in the peripheral blood were determined by cytometric beads array (CBA). The levels of IL-7 and anti-AChR antibody were measured with ELISA. Results No obvious differences were observed in the frequencies of Tfh, Tem, CD8 + T cells and CD4 + T cells, and the ratio of CD4/CD8 in the peripheral blood of MG patients, compared with HCs. MG subjects presented notably decreased frequencies of Tcm and increased frequencies of Tfr compared with HCs. In addition, elevated levels of IL-2, IL-4, IL-12, IL-17 and IFN-γ and lowered levels of IL-7 were observed in the plasma of MG individuals, compared with HCs. No significant correlations were found among the levels of cytokines and frequencies of T cell subsets. No significant changes were found in AChR antibody levels. Conclusion The results suggest a unique spectrum of the memory T cells and follicular T cells, together with a unique cytokine profile in the MG individuals during treatment.