异芒果苷抑制人骨肉瘤U2OS细胞的恶性生物学行为

Isomangiferin Inhibits the Malignant Biological Behavior of Human Osteosarcoma U2OS Cells

本研究以异芒果苷对人骨肉瘤(U2OS)细胞的恶性生物学行为影响及作用机制进行探索。在U2OS细胞中加入不同浓度梯度的异芒果苷,然后通过MTT法分析细胞迁移和侵袭试验分别研究异芒果苷对U2OS细胞增殖、迁移和侵袭能力的作用。分别通过平板克隆形成和微球体法检测异芒果苷对U2OS细胞克隆形成能力和自我更新能力的影响。利用Western blotting检测异芒果苷对U2OS细胞c-Src蛋白表达的影响,并通过RNA激活技术(RNAa)评估异芒果苷是否通过c-Src蛋白抑制骨肉瘤。结果表明,异芒果苷能够抑制U2OS细胞的增殖并呈剂量依赖性,同时不同浓度异芒果苷对U2OS细胞的克隆形成、迁移和侵袭有抑制作用,并呈现一定的剂量依赖关系,而且异芒果苷能够抑制U2OS癌干细胞自我更新的能力。另外,异芒果苷可下调U2OS细胞的c-Src蛋白的表达。异芒果苷可能是通过降低c-Src的表达实现U2OS细胞的恶性生物学行为,而且在一定程度上呈现剂量依赖性。

To study the inhibitory effect and mechanism of isomangiferin on malignant biological behavior of U2OS cells.Osteosarcoma cell line U2OS was cultured in a normal culture condition and incubated with different concentration of isomangiferin.MTT assay was used to assess the proliferation ability.Transwell small chamber assay was applied to assess the migration and invasion ability.Plate colony formation test was used to analysis the colony formation ability.Serum-free suspension culture was performed to test the microsphere formation ability which reflects self-renewal ability of cancer stem cells.Western blotting was applied to analysis the change of pro-oncogene c-Src after isomangiferin treatment.RNA activation(RNAa)technology was applied to assess whether c-Src mediated the inhibitory action of isomangiferin on osteosarcoma.MTT assay suggested that isomangiferin inhibited proliferation ability of U2OS dose-dependently.Plate colony formation and transwell small chamber assay revealed that isomangiferin suppressed the colony formation,migration and invasion ability of U2OS dose-dependently.Microsphere formation assay suggested that isomangiferin may inhibit the self-renewal ability of U2O cancer stem cells.c-Src protein could be downregulated by isomangiferin in U2OS and c-Src RNAa can partly reverse the inhibitory effect of isomangiferin on osteosarcoma.Isomangiferin can compromise the malignant biological behavior and stemness of osteosarcom in dose-dependent manner.Moreover,this inhibition effect may be by downregulation c-Src expression.