山东医药2017，Vol.57Issue(18)：16-19,4.DOI:10.3969/j.issn.1002-266X.2017.18.005
SGK-1在缺氧诱导小鼠垂体瘤细胞AtT-20增殖、凋亡中的表达及作用探讨
Expression and role of SGK-1 in hypoxia-induced proliferation and apoptosis of mouse pituitary tumor cells AtT-20
摘要
Abstract
Objective To observe the expression of serum and glucocorticoid-regulated kinase-1 (SGK-1) in mouse pituitary tumor cells AtT-20′s proliferation and apoptosis induced by hypoxia and to explore the role of SGK-1 in the process.Methods We classified AtT-20 cells into the observation group and control group randomly, then put 50, 100, 200 μmol/L CoCl2 (for modulating the hypoxia) in the observation group while nothing was put in the control group, next, we tested the A570 value of cell proliferation at 0, 24, 48, 72, 96 h by MTT.We added 200 μmol/L CoCl2 to AtT-20 cells, when the cells were cultured for 0, 24, 48, 72, 96 h, we calculated the apoptosis rate by flow cytometry, separately.Meanwhile, we detected the mRNA and protein of SGK-1 by semi quantitative PCR and Western blot.After that, we divided AtT-20 cells into 4 groups, cells in the groups A and B were transfected with interfering plasmid SGK-1 siRNA, and cells in the groups C and D were transfected with control interfering plasmid siCon.At 24 h after transfection, cells in the groups A and C were added with 200 μmol/L CoCl2, while cells in the groups B and D were not treated.Finally, we tested the A570 value and the apoptosis rate at 0, 24, 48, 72, 96 h by MTT and flow cytometry, respectively.Results At 72 h after hypoxia, A570 value of AtT-20 cells in the observation group decreased gradually with the increasing concentration and prolonged time, and difference was statistical significant as compared with that of the control group (all P<0.05).The apoptosis rate had no obvious change at 24, 48 h, and the apoptosis rate increased at 72, 96 h as compared with that at 0 h (all P<0.05).The mRNA and protein expression of SGK-1 increased at 24, 48 h (all P<0.05), but had no change at 72 h as compared with that at 0 h (all P>0.05).The A570 value in the group A at every time point after hypoxia was lower than that of the other groups, while the A570 value in the group C at 72, 96 h was higher than that of the group A and lower than that of groups B, C, meanwhile, significant difference was found between them (all P<0.05).The apoptosis rate in the group A at every time point after hypoxia was higher than that of the other groups, while the apoptosis rate in the group C at 72 h was lower than that of the group A and higher than that of groups B, C, meanwhile, significant difference was found between them (all P<0.05).Conclusion Hypoxia inhibits proliferation of AtT-20 cells and promotes its apoptosis, and the increasing expression of SGK-1 in the process protects cells from damage induced by hypoxia.关键词
脑缺氧/垂体瘤细胞/血清和糖皮质激素调节蛋白激酶/氯化钴/基因干扰/细胞凋亡/细胞增殖Key words
cerebral hypoxia/pituitary tumor cell/serum and glucocorticoid-regulated kinase (SGK)/CoCl2/gene interference/cell apoptosis/cell proliferation分类
医药卫生引用本文复制引用
韦睿,吴杰,曾伟,张玮..SGK-1在缺氧诱导小鼠垂体瘤细胞AtT-20增殖、凋亡中的表达及作用探讨[J].山东医药,2017,57(18):16-19,4.基金项目
国家自然科学基金资助项目(81560319) （81560319）
云南省应用基础研究面上项目(2016FB130) （2016FB130）
云南省卫生科技计划项目 (2016NS066). （2016NS066）
