Bondza Patrick Kibangou, Metz Christine N, Akoum Ali
Laboratoire d'Endocrinologie de la Reproduction, Centre de Recherche, Hôpital Saint-François d'Assise, Centre Hospitalier Universitaire de Québec, Faculté de Médecine, Université Laval, 10 rue de l'Espinay, Local D0-711, Québec G1L 3L5, Canada.
J Reprod Immunol. 2008 Apr;77(2):142-51. doi: 10.1016/j.jri.2007.07.004. Epub 2007 Sep 12.
Human endometrium undergoes a series of dynamic physiological changes during the menstrual cycle of reproductive age women. Many factors, including hormones, cytokines, growth factors, matrix metalloproteinases and integrins, are essential for the success of embryonic implantation into endometrial tissue. Herein, we used a well-differentiated endometrial adenocarcinoma cell line, Ishikawa, to investigate in vitro the role played by macrophage migration inhibitory factor (MIF) in the regulation of endometrial receptivity markers. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that MIF induced a slight increase in alpha(v) (alphav) mRNA integrin subunit expression during the first 12h, but reached a significant difference after 24h MIF treatment compared to control, whereas beta(3) (beta3) integrin subunit displayed significant increase in mRNA 2h following treatment. Immunocytofluorescence showed strong alphav and beta3 immunostaining at 25 ng/ml MIF, and Western blotting clearly indicated increased alphav and beta3 protein expression. MIF treatment significantly stimulated vascular endothelial growth factor (VEGF) mRNA expression in a dose- and time-dependent manner after 24 h treatment. Moreover, immunocytofluorescence revealed positive VEGF immunostaining compared to control, and analysis by ELISA of VEGF release in culture supernatants demonstrated that MIF (25 ng/ml) significantly induced VEGF secretion at 12 and 24 h. In conclusion, this study provides evidence that MIF directly up-regulates alphavbeta3 integrin and VEGF expression in human endometrial Ishikawa cells and may advance our understanding of factors involved in the establishment of endometrial receptivity and successful implantation.
在育龄期女性的月经周期中，人类子宫内膜会经历一系列动态的生理变化。许多因素，包括激素、细胞因子、生长因子、基质金属蛋白酶和整合素，对于胚胎成功植入子宫内膜组织至关重要。在此，我们使用一种高度分化的子宫内膜腺癌细胞系 Ishikawa，在体外研究巨噬细胞迁移抑制因子（MIF）在调节子宫内膜容受性标志物中所起的作用。定量实时聚合酶链反应（qRT-PCR）显示，MIF 在最初 12 小时内诱导α(v)（αv）mRNA 整合素亚基表达略有增加，但与对照组相比，在 MIF 处理 24 小时后达到显著差异，而β(3)（β3）整合素亚基在处理后 2 小时 mRNA 显示出显著增加。免疫细胞荧光显示在 25 ng/ml MIF 时αv 和β3 免疫染色强烈，蛋白质印迹清楚地表明αv 和β3 蛋白表达增加。MIF 处理 24 小时后，以剂量和时间依赖性方式显著刺激血管内皮生长因子（VEGF）mRNA 表达。此外，免疫细胞荧光显示与对照组相比 VEGF 免疫染色呈阳性，通过 ELISA 分析培养上清液中 VEGF 的释放表明，MIF（25 ng/ml）在 12 小时和 24 小时显著诱导 VEGF 分泌。总之，本研究提供了证据表明 MIF 直接上调人子宫内膜 Ishikawa 细胞中αvβ3 整合素和 VEGF 的表达，并可能增进我们对参与子宫内膜容受性建立和成功植入的因素的理解。
