DFU对骨肉瘤细胞增殖和侵袭能力的影响

The inhibitory effects of DFU on proliferation and invasion of osteosarcoma cells

目的观察选择性环氧合酶-2抑制剂DFU对人骨肉瘤细胞株MG-63增殖和侵袭能力的影响及其对Ang-2基因表达的调控。方法体外培养骨肉瘤细胞株MG-63，免疫荧光法检测Ang-2在MG-63细胞中的表达，应用噻唑蓝（MTT）比色法和形态学检测研究不同浓度DFU对骨肉瘤细胞株MG-63的生长抑制作用，Boyden小室体外侵袭实验检测其对骨肉瘤细胞侵袭能力的影响，RT—PCR法分析DFU对骨肉瘤细胞株MG-63中Ang-2基因表达的影响。结果免疫荧光染色结果显示Ang-2在骨肉瘤细胞株MG-63中呈阳性染色，MTT法和形态学检测提示DFU可抑制骨肉瘤细胞株MG-63的增殖，并具有剂量依赖性，在DFU浓度为200μmol／L时，对细胞生长有明显的抑制作用。Boyden小室体外侵袭实验显示DFU可降低骨肉瘤细胞的侵袭能力，在DFU浓度为200μmol／L时，作用最显著。逆转录-聚合酶链反应（RT—PCR）分析显示经DFU作用后的骨肉瘤细胞株MG-63表达Ang-2较用药前明显下降。结论DFU对骨肉瘤细胞株MG-63的增殖有抑制作用，其可能通过抑制Ang-2的表达，降低其侵袭能力。

Objective To investigate the inhibitory effects of DFU, a Cyclooxygenase-2-selectedinhibitor, on proliferation and invasion of osteosarcoma cells line MG-63, and the influences of DFU on the expression of angiopoietin-2 （Ang-2） in MG-63 cells. Methods Osteosarcoma cells line MG-63 was cultured in vitro. The expression of Ang-2 in MG-63 was observed by immunofluorescence. Methyl thiazolyl tetrazolium （MTT） and morphology were used to study the inhibitory effects of DFU with different concentration on osteosarcoma cells line MG-63. Boyden chamber in vitro invasion assay was applied to observe the changes of invasion capability. The expression of Ang-2 of osteosarcoma cells line MG-63 was analysed by RT-PCR. Results Positive staining was seen in the cells line MG-63 by immunofluorescence. The inhibition rate of osteosarcoma cells line MG-63 was higher when treated with DFU and the rate reached the peak when the concentration of DFU was 200 μmol/L. The invasion ability was remarkably decreased after treated with DFU. Ang-2 mRNA expression of osteosarcoma cells line MG-63 declined obviously than before medication. Conclusion DFU has lethal effect on osteosarcoma cells line MG-63. It may inhibit the proliferation and invasion of osteosarcoma cells line MG-63 by inhibiting the expression of Ang-2.