ELK1调控PI3K/Akt信号通路促进骨肉瘤细胞增殖及其机制研究

Function and mechanism of ELK1 regulating PI3K/Akt signaling pathway to promote osteosarcoma proliferation

摘要:

目的 探讨E26转录因子1（ELK1）调控PI3K/Akt信号通路对骨肉瘤细胞增殖能力的影响及其发挥作用的可能机制。方法 Western blotting检测ELK1在骨肉瘤细胞系U2OS、MG-63、HOS及正常成骨细胞hFOB11.9中的相对表达量；选择相对表达量最高的骨肉瘤细胞系进行ELK1 siRNA转染，分为si-NC组、si-ELK1-1组和si-ELK1-2组；MTS检测各组细胞增殖率；平板克隆实验检测各组细胞克隆形成能力；Western blotting检测各组细胞中PI3K/Akt信号通路关键蛋白pPI3K和pAKT的相对表达量；采用PI3K/Akt信号通路激活剂SC79与ELK1 siRNA同时处理骨肉瘤细胞后MTS法检测各组细胞增殖率，平板克隆实验检测各组细胞克隆形成能力。结果 与正常成骨细胞比较，ELK1在骨肉瘤细胞系中的相对表达量均升高（P <0.05），在U2OS细胞中的相对表达量最高（P <0.05）；U2OS细胞转染ELK1 siRNA，Western blotting结果显示，与si-NC组比较，si-ELK1-1组、si-ELK1-2组U2OS细胞中ELK1的相对表达量降低（P <0.05）；与si-NC组比较，si-ELK1-1组和si-ELK1-2组U2OS细胞增殖率和克隆形成能力均降低（P <0.05），细胞中pPI3K和pAkt蛋白的相对表达量降低（P <0.05）。SC79处理si-ELK1组U2OS细胞后，ELK1 siRNA对细胞增殖和克隆形成能力的抑制作用减弱（P <0.05）。结论 ELK1在骨肉瘤细胞中高表达，通过激活PI3K/Akt信号通路促进骨肉瘤细胞增殖的能力，ELK1可作为治疗骨肉瘤的潜在靶点。

Abstract:

Objective To study the effect of E26 transcription factor 1 (E-twenty six transcription factor 1, ELK1) on the proliferation of osteosarcoma cells by regulating the PI3K/Akt signaling pathway, and to explore the possible mechanism of its role.Methods Western blotting was used to detect the expression level of ELK1 in osteosarcoma cell lines MG-63, HOS, U2OS, and normal osteoblast hFOB11.9. High-expressing osteosarcoma cell line was selected for ELK1 siRNA infection and divided into si-NC group, si-ELK1-1 group, and si-ELK1-2 group. Western blotting was used to detect the expression level of ELK1 in each group of cells; MTS was used to detect the cell proliferation rate of each group; plate cloning experiment was used to detect the cell formation ability of each group; western blotting was used to detect the effects of PI3K/Akt signaling pathway-related proteins in each group of cells. PI3K/Akt signaling pathway activator SC79 and ELK1 siRNA were used to treat osteosarcoma cells at the same time. MTS was used to detect the cell proliferation rate of each group. Plate cloning experiment was used to detect the cell clone formation ability of each group.Results Western blotting results showed that the expression of ELK1 in osteosarcoma cell lines was higher than that in normal osteoblasts (P < 0.05), and the expression was highest in U2OS cells (P < 0.05). U2OS cells were transfected with ELK1 siRNA. Western blotting showed that compared with the NC group, the expression of ELK1 in si-ELK1-1 and si-ELK1-2 groups U2OS cells were reduced (P < 0.05); compared with the si-NC group, the proliferation rate and cloning ability of si-ELK1-1 and si-ELK1-2 U2OS cells were decreased (P < 0.05), and the expression of pPI3K and pAkt proteins were decreased (P < 0.05). After SC79 treatment of U2OS cells in the si-ELK1 group, the inhibitory effect of ELK1 siRNA on cell proliferation and clone formation was weakened (P < 0.05).Conclusion Interfering with the expression of ELK1 can inhibit the proliferation of osteosarcoma cells by regulating PI3K/Akt.