缺氧对细胞滋养细胞MMP9/TIMP1基因表达及细胞侵袭力的影响

Effects of Hypoxia on Expression of Matrix Metalloproteinase 9/Tissue Inhibitor of Metalloproteinase 1 and Invasion Ability of Cytotrophoblasts in vitro

目的探讨缺氧对细胞滋养细胞基质金属蛋白酶9(MMP9)及其抑制物1(TIMP1)mRNA和蛋白的表达及细胞侵袭能力的影响。方法将永生化的早孕细胞滋养细胞进行化学缺氧(150μmol/L CoCl2),用免疫组化、荧光定量PCR、Western blot等方法检测正常和缺氧细胞滋养细胞系MMP9/TIMP1基因的表达,应用Transwell侵袭模型检测缺氧对细胞滋养细胞侵袭能力的影响。结果缺氧组的细胞滋养细胞MMP9、TIMP1 mRNA及蛋白表达均升高(P<0.05),MMP9/TIMP1比值升高,且呈时间依赖性。缺氧组细胞滋养细胞的侵袭力较对照组增强(P<0.05)。结论在生理性缺氧条件下,早孕细胞滋养细胞的侵袭力是增加的,并与MMP9/TIMP1比值的升高有关;同时TIMP1除了调节MMP9外,还可能促进细胞滋养细胞的增殖与分化。

Objective To investigate the effects of hypoxia on the expression of matrix metalloproteinase 9（MMP9）/tissue inhibitor of metalloproteinase 1（TIMP1） and invasion ability of cytotrophoblasts in vitro.Methods Immortalized human extravillous trophoblast cell line was induced under hypoxia circumstance by 150 μmol/L CoCl2.Immunocytochemistry,fluorescence quantitative real-time PCR and Western blot were used to determine the gene expression of MMP9/TIMP1 in cytotrophoblasts.The invasion of hypoxia-induced cytotrophoblasts was assessed by Transwell chamber assay.Results The mRNA and protein expression levels of MMP9 and TIMP1 were up-regulated in hypoxia group（P0.05）,and the ratio of MMP9/TIMP1 was increased with prolonged time.The invasion ability in hypoxia group was significantly enhanced as compared with control group（P0.05）.Conclusion Physiological hypoxia could enhance the invasion ability of cytotrophoblasts during early pregnancy with the increase of the ratio of MMP9/TIMP1,and TIMP1 may promote cytotrophoblast proliferation and differentiation besides regulating MMP9.