美洛昔康对结肠癌细胞VEGF和angiopoietin-2表达的影响

Effects of meloxicam on vascular endothelial growth factor and angiopoietin-2 expression in colon carcinoma cell line HT-29

目的:研究COX-2选择性抑制剂美洛昔康(meloxicam)对结肠癌细胞HT-29生长及血管内皮生长因子(VEGF)和血管生成素-2(angiopoietin-2,Ang-2)表达的影响.方法:分别用100,200,400,800μmol/L美洛昔康对细胞进行干预后,采用CCK-8活细胞计数法检测细胞增殖,流式细胞术检测细胞周期,ELISA测定细胞培养上清液中VEGF,Ang-2的蛋白含量,实时荧光定量PCR检测细胞COX-2,VEGF,Ang-2mRNA含量.结果:美洛昔康作用不同时间后,对HT-29细胞具有细胞毒作用,细胞增殖活性随浓度增加、时间延长逐渐降低(P值:0.000→0.029;0.000→0.043),呈现量-效、时-效关系.并且美洛昔康呈浓度依赖性改变细胞周期分布,G0/G1期细胞比例增加(P值:0.000→0.015).上清液中VEGF和Ang-2蛋白含量明显降低,存在时间和浓度依赖性(P值:0.000→0.018;0.000→0.028).细胞COX-2,VEGF和Ang-2mRNA表达亦明显降低(P值:0.000→0.025),也存在浓度依赖性.结论:美洛昔康能够在蛋白、核酸水平上抑制结肠癌细胞分泌VEGF和Ang-2,从而抑制肿瘤血管生成.

AIM： To investigate the effect of meloxicam, a nonsteroidal anti-inflammatory drug （NSAID）, on the expression of vascular endothelial growth factor （VEGF） and angiopointin-2 （Ang-2） in colon carcinoma cell line HT-29. METHODS： The cultured HT-29 cells were treated with different concentrations of meloxicam （100, 200, 400, 800 μmol/L） for different time. The proliferation of HT-29 cells was detected by Cell Counting Kit 8 （CCK8）. Cell cycle was detected by flow cytometry. The levels of VEGF and Ang-2 protein in the supernatants were determined by enzyme-linked immunosorbent assay （ELISA） and the expression of VEGF and Ang-2 mRNA in HT-29 cells were examined by real-time quantitative reverse-transcriptionpolymerase chain reaction （RT-PCR）. RESULTS： Meloxicam inhibited the proliferation of HT-29 cells in a concentration- and timedependent manner （P = 0.000→0.029; 0.000→0.043）, and the proliferating activity was decreased with the increase of concentration and prolonging of action time. Meloxicam changed the cycles of HT-29 cells in a concentration-dependent manner, and the number of cells at G0/ G1 phase was significantly increased （P = 0.000→0.015）. The levels of VEGF and Ang-2 protein in supernatants were reduced gradually with the increase of concentration or action time （P = 0.000→0.018; 0.000→0.028）. The expression of COX-2, VEGF and Ang-2 mRNA were also reduced with the increase of Meloxicam concentration （P = 0.000→0.025）, CONCLUSION： Meloxicam can reduce the expression of VEGF and Ang-2 at the levels of protein and mRNA in colon carcinoma cell line.