黄芪、莪术配伍对胃癌MKN-45细胞COX-1、COX-2、NF-κB、VEGF、MMP-2表达的影响

Influence of compatibility of Astragalus mongholicus and blue turmeric rhizome on expression of COX-1,COX-2,NF-κB,VEGF and MMP-2 in gastric cancer cell MKN-45

目的观察黄芪、莪术配伍对胃癌MKN-45细胞VEGF、COX-1、COX-2、NF-κB、MMP-2表达的影响。方法以黄芪、莪术配伍作用于MKN-45细胞,并设塞莱希布组、黄芪组、莪术组和空白组相对照。通过ELISA法检测VEGF的水平变化,用RT-PCR以及Western blot方法检测加药后人胃癌细胞COX-1、COX-2、NF-κB、MMP-2的mRNA及COX-2蛋白表达情况。结果各药物组对VEGF表达的抑制效应起始于48 h,配伍组作用大于黄芪组和莪术组。各药物对COX-2 mRNA、NF-κB mRNA和MMP-2 mR-NA均有抑制作用。对COX-2的抑制以塞莱希布组和配伍组最明显,且呈时间依赖性。对NF-κB mRNA的抑制作用起始于48 h,配伍组在作用72 h后对其抑制作用最显著。对MMP-2的抑制作用以塞莱希布组和配伍组较明显,72 h时的作用最强。Western blot曝光结果可见,各组细胞在72kD处可见特异性的COX-2蛋白表达条带。进行灰度分析发现:各药物组对COX-2的抑制作用呈时间依赖性,以中药配伍和塞莱希布的抑制作用最强。结论黄芪、莪术配伍可能是通过对COX-2和NF-κB的调控影响MMP-2、VEGF的表达,从而抑制肿瘤细胞的浸润、转移。

Objective It is to observe the influence of compatibility of Astragalus mongholicus and blue turmeric rhizome on the expression of COX - 1, COX - 2, NF - κB, VEGF and MMP - 2 in gastric cancer cell MKN - 45. Methods The compatibility of Astragalus mongholicus and blue turmeric rhizome was used on MKN - 45 cell, comparing with another five contrasting medicine including celecoxib, Astragalus mongholicus, blue turmeric rhizome and blank drug. Experssion of VEGF was detecded by ELISA. mRNA of COX - 1, COX - 2 , NF - κB, MMP - 2 and COX - 2 proteinum were determined through RT - PCT and Western blot methods. Results Suppression of all medicines on the expression of VEGF began after 48 h. The effect of compatibility was better than that of Astragalus mongholicus and blue turmeric rhizome. The influence of every medicine on COX - 2 mRNA, NF - κB mRNA and MMP - 2 mRNA was obvious. The suppression of celecoxib and compatibility on COX - 2 were both well, and became more apparent following time. All medicines had suppressed expression of NF- κB mRNA after 48 h, and the suppression of compatibility were most apparent after 72 h. The suppression of celecoxib and compatibility was visible and became most apparent after 72 h. Western blot results showed that a specific protein strip of COX - 2 expression was observed at the 72 kD. Gray scale analysis showed that all medicine had obvious suppression on COX - 2, and become more obviously following time. The suppression of celecoxib and compatibility was the strongest. Conclusion The compatibility of Astragalus mongholicus and blue turmeric rhizome has effect on the expression of MMP - 2 and VEGF by regulating COX - 2 and NF - κB, so to inhibit infiltration and metastasis of tumor cell.