马钱子碱抑制骨肉瘤的作用机制

Brucine Regulates the PI3K-AKT Signaling Pathway to Inhibit Osteosarcoma

目的探究马钱子碱对骨肉瘤细胞的抑制作用及可能机制。方法在HOS和143B细胞中分别加入不同浓度的马钱子碱处理细胞,使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法,通过EDU实验和细胞克隆形成实验检测肿瘤细胞的增殖活性;利用细胞划痕实验检测细胞的迁移能力,Transwell小室实验检测骨肉瘤细胞的侵袭能力;流式细胞术检测细胞周期及凋亡情况,引入生物信息预测技术预测马钱子碱对骨肉瘤发挥作用的机制,最后采用Western blot实验验证蛋白的表达量。结果马钱子碱呈时间和剂量依赖性抑制HOS和143B细胞的增殖,也能够抑制肿瘤细胞的迁移和侵袭;马钱子碱通过抑制PI3K-AKT信号通路促进骨肉瘤HOS和143B细胞的凋亡,下调了HOS和143B细胞中p-PI3K、p-AKT、MMP9、CyclinD1、C-caspase9蛋白含量。结论马钱子碱可呈剂量依赖性的抑制HOS和143B细胞的增殖和侵袭迁移,有望作为潜在的抗骨肉瘤药物。

Objective To investigate the inhibitory effect of brucine on osteosarcoma cells and its possible mechanism.Methods HOS and 143B cells were treated with brucine at different concentrations.3-(4,5-dimethylthiazol-2)-25-diphenyltetrazolium bromide(MTT)colorimetry,EDU assay and cell clonal formation assay were used to detect the proliferative activity of tumor cells.The migration ability of osteosarcoma cells was detected by cell scratch assay,and the invasion ability of osteosarcoma cells was detected by Transwell cell assay.Flow cytometry was used to detect cell cycle and apoptosis.Bioinformation prediction technology was introduced to predict the mechanism of brucine on osteosarcoma,and finally Western blot assay was used to verify the protein expression level.Results Brucine inhibited the proliferation of HOS and 143B cells in a time-dependent and dose-dependent manner,and also inhibited the migration and invasion of tumor cells.Brucine promoted the apoptosis of osteosarcoma HOS and 143B cells by inhibiting the PI3K-AKT signaling pathway,and down-regulated the contents of p-PI3K,p-AKT MMP9,CyclinD1 and C-caspase9 proteins in HOS and 143B cells.Conclusion Brucine can inhibit the proliferation,invasion and migration of HOS and 143B cells in a dose-dependent manner which is expected to be a potential anti-osteosarcoma drug.