磷脂酰肌醇3激酶/蛋白激酶B信号通路对MC3T3-E1细胞增殖和周期调控的研究  

Effects of PI3K /Akt signaling pathway on the proliferation and cell cycles of MC3T3-E1 cells

作　　者：范文斌[1] 赵建宁[1] 包倪荣[1] 

机构地区：[1]南京大学医学院临床学院(南京军区南京总医院)骨科,南京医学硕士研究生210002

出　　处：《医学研究生学报》2014年第4期347-351,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81000814)

摘　　要：目的近年的研究显示,磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kenase B,PKB/Akt)信号通路是骨形成和骨重建的关键调控信号,文中研究抑制PI3K/Akt信号通路后对MC3T3-E1细胞增殖和细胞周期的影响。方法用PI3K/Akt信号抑制剂PF-04691502处理MC3T3-E1细胞后,通过MTS法检测细胞增殖的活性,流式细胞仪检测细胞周期的变化,Western blot检测细胞周期相关蛋白的表达。结果应用抑制剂抑制PI3K/Akt信号通路后,与对照组细胞存活率(100±5.6)%比较,30、150、750 nmol/L的MC3T3-E1细胞增殖活性明显降低[(64.7±4.1)%、(42.3±1.1)%、(15.9±0.4)%,P<0.01],且随着药物浓度的增加,细胞增殖活性降低越明显。同时,与对照组比较,1μmol/L PF-04691502处理组sub-G1期细胞比例明显升高[(1.45±0.43)vs(0.27±0.21),P<0.01],且细胞周期蛋白D1表达降低。结论抑制PI3K/Akt信号通路能将MC3T3-E1细胞阻滞于G1期,能降低细胞的增殖能力。Objective Recent studies show that the PI3K/Akt signaling pathway is a key regulatory signal in both bone for-mation and bone remodeling .The experiment aimed to investigate the effects of inhibiting the PI 3K/Akt signaling pathway on the prolif-eration and cell cycles of MC3T3-E1 cells. Methods We treated MC3T3 -E1 cells with PF-04691502 the inhibitor of the PI3K/Akt signaling pathway .Then we determined the proliferative activity of the cells by MTS assay , detected the cell cycles by flow cytometry , and measured the expression of cell cycle-related proteins by Western blot . Results After inhibition of the PI3K/Akt signaling path-way with PF-04691502 at 30, 150 and 750 nmol/L, the proliferation of MC3T3-E1 cells was reduced in a dose-dependent manner to 0.647 ±0.041, 0.423 ±0.011 and 0.159 ±0.004, respectively, as compared with 1 ±0.056 in the control group （P＆lt;0.01）.Sim-ultaneously , the percentage of the cells in the sub-G1 phase was significantly higher in the 1 μmol/L group than in the control （[1.45 ±0.43]%vs [0.27 ±0.21]%, P＆lt;0.01） and the expression of cyclin D1 was decreased. Conclusion Inhibition of the PI3K/Akt signaling pathway can block MC 3T3-E1 cells in the sub-G1 phase and reduce the proliferation of the cells .

关 键 词：成骨细胞 磷脂酰肌醇3激酶 蛋白激酶B信号通路 细胞周期 增殖活性 

分 类 号：Q28[生物学—细胞生物学]