下调Notch3表达对骨肉瘤细胞生物学特性的影响

目的 观察Notch3在骨肉瘤细胞中的表达及下调Notch3表达后对骨肉瘤细胞增殖、凋亡及侵袭迁移的影响,探讨Notch3在骨肉瘤细胞生物学功能中的作用.方法 体外培养人骨肉瘤细胞MG63、Saos2、U20S,采用Real-time PCR和Western blot检测3种骨肉瘤细胞系中Notch3的mRNA和蛋白表达水平;利用siRNA敲低Notch3在骨肉瘤U20S细胞中的表达,并将细胞分为siNotch3组和阴性对照组,检测下调Notch3的表达后骨肉瘤细胞增殖、凋亡及侵袭、迁移等生物学特性的变化.结果 Notch3在骨肉瘤细胞MG63、Saos2、U20S中均呈阳性表达,并且在低级别骨肉瘤细胞系MG63中的表达明显低于高级别细胞系Saos2和U20S(P ＜0.05),下调Notch3的U20S细胞(siNotch3组)增殖、侵袭、迁移能力较阴性对照组减弱,凋亡率增加,差异均有统计学意义(P＜0.05).结论 在骨肉瘤细胞U20S中下调Notch3的表达可明显抑制细胞的增殖、侵袭、迁移能力,促进凋亡.Notch3可能是Notch 通路在骨肉瘤中的关键受体之一,并参与了骨肉瘤的发生、发展.

Effect of down-regulating Notch3 expression on biological characteristics of osteosarcoma cells

Objective To explore the expression of Notch3 in human of osteosarcoma cells and determine the effect of its down-regulation on the proliferation,apoptosis,invasion and migration,and to investigate its role in the biological function of the cells.Methods Real-time PCR and Western blotting were used to examine the expression of Notch3 at mRNA and protein levels in osteosarcoma cell lines.siRNA was used to knockdown the expression of Notch3 in U2OS cells.Then the proliferation,apoptosis and metastasis of the cells after transfection were detected for the effects of Notch3 down-regulation.Results Notch3 was highly expressed in human osteosarcoma cell lines MG63,Saos2 and U2OS,but its level was significantly lower in MG63 cells derived from low-grade osteosarcoma than those in Saos2 and U2OS cells derived from high-grade osteosarcoma (P ＜ 0.05).When compared with untreated cells (Control),down-regulation of Notch3 in U2OS cells markedly repressed cell proliferation,decreased invasion and migration (P ＜ 0.05).but enhanced apoptosis (P ＜ 0.05).Conclusion Down-regulation of Notch3 reduces the proliferation,invasion,migration,but increases the apoptosis in osteosarcoma cells.Notch3 may be a key receptor of Notch pathway involving in the occurrence and development of osteosarcoma.