雌、孕激素和肝素结合生长因子对Ishikawa细胞金属蛋白酶组织抑制物-1表达的调节

Regulatory effect of estrogen, progestin and HB-EGF on the expression of tissue inhibitor of metalloproteinase 1 in Ishikawa cells

目的通过体外实验研究雌激素、孕激素、肝素结合生长因子(HB-EGF)对Ishikawa细胞中基质金属蛋白酶-9 (MMP-9)、金属蛋白酶组织抑制物-1(TIMP-1)蛋白表达的调节及其在胚胎种植中的意义。方法体外培养高度分化的子宫内膜癌细胞Ishikawa,按实验目的分别加入雌激素、孕激素、雌孕激素联合、雌孕激素与米非司酮(RU486)联合、HB-EGF刺激48 h后,采用免疫细胞化学、Western blot两种方法测定各种条件下Ishikawa细胞中MMP-9、TIMP-1蛋白的表达。结果雌、孕激素单独或联合作用48 h后都可以显著降低Ishikawa细胞中TIMP-1蛋白的表达(P<0．05),同时加RU486后 TIMP-1蛋白的下降趋势减弱。相反,加HB-EGF刺激48 h后TIMP-1蛋白的表达增高(P<0．05)。各种因素处理48 h后, Ishikawa细胞中MMP-9蛋白均没有阳性表达。结论雌、孕激素对TIMP-1蛋白的表达具有下调作用,RU486可抑制孕激素的下调作用;HB-EGF对TIMP-1蛋白的表达具有上调作用;MMP-9蛋白在Ishikawa细胞中无表达。

Objective： To investigate the regulatory effect of estrogen, progestin and heparin-binding epidermal growth factor（HB-EGF） on the matrix metalloproteinase （MMP） 9 and the tissue inhibitor of metalloproteinase（TIMP） 1 in Ishikawa cells. Methods： The highly differentiated endometrial adenocarcinoma cells （Ishikawa cell line） were incubated with 17-beta estradiol （1 × 10^-8 mol/L）, medroxyprogesterone acetate （MPA） （1 × 10^-6 mol/L）, RU486 （1 ×10^-8mol/L） or HB-EGF（10 ng/ml） for 48 hours respectively. The expression of MMP-9 protein and TIMP-1 protein were detected by immunocytochemistry and Western blot. Results： Either estrogen alone, progestin alone or progestin combined with estrogen could significantly decreasethe expression of TIMP-1 protein after 48 hours of treatment （P〈0. 05）. But this down-regulation was inhibited by the addition of RU486. On the contrary, HB-EGF elevated the expression level of TIMP -1 protein after 48 hours of treatment （P〈0. 05）. The MMP-9 protein did not be detected in Ishikawa cells. Conclusions. （1） Both estrogen and progestin can down-regulate the expression of TIMP-1 protein in Ishikawa cells, but RU486 can inhibit this effect of down-regulation. （2） HB-EGF can elevate the level of TIMP-1 protein. （3）There is no expression of MMP-9 protein in Ishikawa cells.