缺氧对肺腺癌细胞A549MMP-9、TIMP-1表达的影响

目的:研究缺氧条件下基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶组织抑制因子-1(TIMP-1)的表达变化规律。方法:人肺腺癌细胞A549常氧/缺氧培养,酶谱法测定培养基中MMP-9酶活性,免疫细胞化学、免疫印迹技术分别检测MMP-9及TIMP-1原位蛋白及分泌蛋白表达量。结果:MMP-9、TIMP-1的阳性染色颗粒主要呈胞浆着色。缺氧48h内,MMP-9酶活性、原位蛋白及分泌蛋白表达均无显著变化;随缺氧时间延长,TIMP-1原位蛋白及分泌蛋白表达均有升高趋势(P<0.05)。结论:缺氧24h内MMP-9表达不增加,可能与癌基因、激酶活化引发的细胞筛选有关;TIMP-1在缺氧状态下表达增多,可能与其多种功能相关。

The effect of oxygen deficiency on the expressions of MMP-9 and TIMP-1 in human lung adenocarcinoma A549 cell line

Objective To study the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1) in human lung adenocarcinoma A549 cell line under oxygen deficient conditions. Methods The A549 cell line was cultured under normal or oxygen-deficient conditions. The active form of MMP-9 was measured by gelatin zymography, while intracellular and secretive protein of MMP-9 and TIMP-1 were determined by immunocytochemistry and Western blot, respectively. Results Mostly, the cytoplasm was dyed in those cells positive for MMP-9 and TIMP-1 staining. No significant changes in the enzyme activity and protein level of MMP-9 were detectable within 48 hours of oxygen deficiency whereas the intracellular and secretive proteins of TIMP-1 had an increasing tendency with the elongation of hypoxia. Conclusion No increase in the expression of MMP-9 occurs within 24 hours of oxygen deficiency, which may be the result of cell selection induced by oncogene and kinase activation, while more TIMP-1 are expressed under the hypoxic state, which may be associated with its multiple functions.