肉桂醛对黑素瘤A375细胞凋亡和VEGF MMP-9表达的影响

Effect of Cinnamic Aldehyde on Apoptosis and VEGF, MMP-9 Expression in Human Melanoma A375 Cells

目的研究肉桂醛对体外培养的人黑素瘤细胞株A375凋亡和血管内皮细胞生长因子(VEGF)及基质金属蛋白酶-9(MMP-9)表达的影响。方法 A375细胞在体外二维和三维培养后,在培养基中加入不同浓度(0,10,20和40μmol/L)肉桂醛,24小时后流式细胞仪检测细胞周期和凋亡,Western blot法检测VEGF蛋白水平,明胶酶谱法检测MMP-9分泌情况。结果肉桂醛能促进A375细胞凋亡,降低细胞增殖指数。40μmol/L肉桂醛作用后的A375细胞增殖指数最低(53.38±4.25%vs35.13±3.24%),细胞凋亡百分比最高(4.37±0.25%vs10.50±0.37%),各浓度间作用差异具有统计学意义(P<0.05)。肉桂醛作用后二维和三维培养的人黑素瘤细胞VEGF的表达下降(1.193±0.195vs0.204±0.017;1.549±0.214vs0.392±0.019),MMP-9的分泌也减少(24885.83±865.80vs14150.77±555.63;29408.50±817.62vs15121.43±497.95),各浓度肉桂醛间的抑制强度差异具有统计学意义(P<0.05)。结论肉桂醛能促进人黑素瘤细胞凋亡,抑制其增殖,并通过降低VEGF和MMP-9的表达来抑制人黑素瘤细胞的侵袭,其抑制强度与药物浓度成正比。

Objective To investigate the effect of cinnamic aldehyde on apoptosis and expression of vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （MMP-9） in A375 cells. Methods A375 cells in two-dimensional and three-dimensional culture were treated with cinnamic aldehyde of different concentration （0, 10, 20 and 401xM） for 24 hours. The proliferation and apoptosis of A375 cells was tested by flow cy- tometry. The VEGF protein expression level was detected by Western blot. The level of MMP-9 secretion was detected by gelatin zymography assay. Results Cinnamic aldeyde promoted A375 cells apoptosis and inhibited cell proliferation after 24 hours. The proliferation of A375 ceils was most remarkably inhibited after co-culture of 40lxM cinnamic aldeyde （53.38 ±4.25% vs 35.13 ± 3.24% ）. The ratio of apoptotic cell was most remarkably increased after treated with 401xM cinnamic aldeyde （4.37 -± 0.25% vs 10.50 ± 0.37% ）. The concentration of cinnamic aldeyde affected the cell apoptosis significantly（P 〈0.05）. Cinnamic aldeyde inhibited VEGF expression of A375 ceils in tow-dimensional and three-dimensional culture（ 1. 193 ± 0. 195 vs 0. 204 ±0.017 ;1. 549 ±0.214 vs 0.392 ±0.019）. Cinnamic aldeyde also decreased MMP-9 secretion of A375 cells in tow-dimensional and three-dimensional culture（24 885.83 ± 865.80 vs 14 150.77 ± 555.63; 29 408.50 ± 817.62 vs 15 121.43 ± 497.95）. The concentration of cinnamic aldeyde affected the VEGF and MMP-9 expression significantly （P 〈 0.05）. Conclusion Cinnamic aldeyde could promote the apopto- sis and inhibit the proliferation of human melanoma ceils. Cinnamic aldeyde could prevent the invasion of human melanoma cells by inhibiting VEGF and MMP-9 expression. The efficacy strengthened according to the higher medicine concentration.