S100A6通过PI3K/Akt信号通路促进人骨肉瘤细胞143B增殖和迁移

S100A6 promotes proliferation and migration of human osteosarcoma cell line 143B through PI3K/Akt signaling pathway

目的：研究PI3K/Akt信号通路在S100A6介导的人骨肉瘤细胞143B的增殖和迁移中的作用.方法：首先制备重组的人S100A6蛋白（recombinant human S100A6,rhS100A6）;rhS100A6与PI3K抑制剂（LY294002和wortmannin）单独或同时处理143B细胞,其中rhS100A6的终浓度为30 mg/L,LY294002和wortmannin的终浓度分别为10 μmol/L和0.5 μmol/L;采用Western blotting分析143B细胞中PI3K/Akt信号通路相关分子总Akt（total Akt,t-Akt）及磷酸化Akt（phosphorylation of Akt,p-Akt）蛋白的表达变化,MTT检测细胞增殖,Transwell检测细胞迁移.结果：（1）成功制备rhS100A6蛋白,rhS100A6显著增强143B细胞的增殖和迁移能力（P〈0.05）;（2）rhS100A6上调143B细胞中Akt的磷酸化;（3）与rhS100A6组相比,rhS100A6与LY294002或wortmannin联合处理组143B细胞的p-Akt减少（P〈0.05）,细胞的增殖和迁移能力降低,在不同时点细胞的增殖率下降10.3%～69.7%,细胞迁移率下降37.9%～41.6%,差异均有统计学意义（P〈0.05）.结论：S100A6促进人骨肉瘤细胞143B增殖和迁移的作用至少部分是通过激活PI3K/Akt信号通路实现的.

AIM： To investigate the effect of PI3 K/Akt signaling pathway on S100A6-induced proliferation and migration of human osteosarcoma cell line Ig3B. METHODS： Recombinant human S100A6 protein （rhS100A6） was prepared. The 143B cells were treated with rhS100A6 in the presence or absence of PI3K inhibitor （ LY294002 or wortmannin） exposure. The final concentrations of rhS100A6, LY294002 and wortmannin were 30 mg/L, 10 μmol/L and 0.5 μmol/L, respectively. The expression levels of total Akt （t-Akt） and phosphorylated Akt （p-Akt） in the 143B cells were analyzed by Western blotting. The cell proliferation and migration were determined by MTT and Transwell assays. RESULTS： rhS100A6 protein was successfully prepared, and significantly increased the proliferation and migration of l d3B cells （P 〈 0. 05）. rhS100A6 up-regulated the phosphorylation of Akt in 143B cells （P 〈 0. 05 ）. Compared with rhS100A6 group, the level of p-Akt in 143B cells and the proliferation and migration of the cells were decreased in combined treatment group of rhS100A6 with LY294002 or wortmannin （ P 〈 0. 05 ）, where the proliferation rate at different time points dropped from 10.3% to 69.7% （P 〈0. 05）, and the migration rate dropped from 34.9% to 47.7% （P 〈0.05）. CON-CLUSION： To some extent, S100A6 promotes proliferation and migration of human ostersarcoma cell line 143B through PI3K/Akt signaling pathway.