壳五糖对骨肉瘤细胞抗肿瘤作用的研究

Antitumor effect of the pentamer of chitooligosaccharide on osteosarcoma cells

目的探讨壳五糖对骨肉瘤细胞的抗肿瘤作用及其作用机制。方法使用不同质量浓度的壳寡糖单糖(聚合度为2~6)处理3种骨肉瘤细胞系(MNNG、MG63、U2OS),48 h后采用CCK-8法评估细胞活力及细胞增殖情况,并通过流式细胞技术分析细胞周期及细胞凋亡的变化情况。利用Transwell实验检测细胞的迁移和侵袭能力。采用蛋白质免疫印迹法检测壳五糖处理前后细胞中磷酸肌醇-3-激酶(PI3K)/丝氨酸-苏氨酸蛋白激酶(AKT)信号转导通路中关键蛋白表达水平的变化。结果壳寡糖单糖以剂量依赖性方式抑制骨肉瘤细胞的增殖,其中以壳五糖作用效果最佳,其在3种骨肉瘤细胞中的半数抑制浓度(IC50)分别为1.55 mg/mL(MNNG)、0.84 mg/mL(MG63)、0.76 mg/mL(U2OS)。壳五糖处理3种骨肉瘤细胞后显著抑制这些细胞的增殖和迁移能力,同时影响骨肉瘤细胞周期并促进细胞凋亡。蛋白质免疫印迹法检测显示,PI3K/AKT信号转导通路中的磷酸化PI3K、磷酸化AKT的表达明显下降。结论壳五糖可有效抑制骨肉瘤细胞的增殖、迁移、侵袭能力,其作用机制可能是通过调控PI3K/AKT信号转导通路产生的。

Objective To investigate the antitumor effect of the pentamer of chitooligosaccharide on osteosarcoma cells and its potential mechanism.Methods Osteosarcoma cells(MNNG,MG63,U2OS)were treated with varying concentrations of chitooligosaccharide monosaccharides(DP=2~6)for 48 h to investigate the cell viability and proliferation according to the CCK-8 method.Cell cycle and apoptosis were detected with the flow cytometry analysis.Transwell assay was used to detect cell migration and invasion.Western blot was used to detect the mechanism of chitooligosaccharide monosaccharides treatment.Results Chitooligosaccharide monosaccharides inhibited the proliferation of osteosarcoma cells in a dose-dependent manner.The pentamer of chitooligosaccharide had the strongest antitumor effect,and its IC50 were 1.55 mg/mL,0.84 mg/mL,0.76mg/mL in osteosarcoma cells(MNNG,MG63,U2OS).Generally,treatment with the pentamer of chitooligosaccharide had significant effects on cell cycle arrest,metastasis inhibition,apoptosis in osteosarcoma cells.Meanwhile,the expression of p-PI3K,p-AKT were significantly decreased.Conclusion The pentamer of chitooligosaccharide can effectively inhibit the proliferation,migration and invasion of osteosarcoma cells,probably through the PI3K/AKT pathway.