Effect of cytoglobin low expression on proliferation of glioma cells and its mechanism / 中华神经医学杂志

ABSTRACT

Objective To explore the effect of cytoglobin (CYGB) low expression on the proliferation of glioma cells and its mechanism.Methods Glioma samples were chosen from patients accepted glioma resection in our hospital from June 2012 to December 2015;primary glioma cells extracted from these samples were cultured and performed purity identification.The nominated cells were divided into transfection of 24 h group,transfection of 48 h group,transfection of 72 h group,and negative control group;cells,excepted from negative control group,were transfected by CYGB shRNA for 24,48 and 72 h,respectively,to inhibit the CYGB expression.CCK-8 assay was used to observe the proliferation of glioma cells after different transfection times (0,1,2,3,4 and 5 d after cell culture).The expressions of CYGB,phosphatidylinositol 3 kinase (PI3K),total alanine aminotransferase (Akt) and phosphorylated (p)-Akt were detected by Western blotting,and the levels of interleukin (IL)-6,IL-10,tumor necrosis factor (TNF)-α,transforming growth factor (TGF)-β and vascular endothelial growth factor (VEGF) were examined by ELISA.Results (1) The proliferation of glioma cells was enhanced at different times after CYGB shRNA transfection,and the optical density showed significant differences at different times after CYGB shRNA transfection (P＜0.05);as compared with those in the negative control group,the cell proliferative capacity and optical density in the transfection of 24 h group,transfection of 48 h group and transfection of 72 h group were significantly increased (P＜0.05).As compared with those in the negative control group,the CYGB protein expression was significantly d ecreased and PI3K and p-Akt protein expressions were significantly increased in the transfection of 24 h group,transfection of 48 h group and transfection of 72 h group,accordingly (P＜0.05),while no significant difference was noted in the total Akt protein expression (P＞0.05).The levels of IL6,IL10,TNF-α,TGF-β,and VEGF were successively increased in the transfection of 24 h group,transfection of 48 h group and transfection of 72 h group as compared with those in the negative control group (P＜0.05).Spearman correlation analysis showed that the expression of CYGB in the glioma was negatively correlated with PI3K and p-Akt expressions,and IL-6,IL-10,TNF-α,TGF-β,and VEGF levels (P＜0.05).Conclusion The effect of cytoglobin on proliferation of glioma cells may be related to the signal pathway of PI3K-Akt.