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目的:了解在缺氧条件下,单层贴壁培养的滋养细胞系TEV-1中,E-钙黏蛋白(E-cadherin,E-cad)的表达在mRNA水平和蛋白水平变化情况,探讨缺氧环境对滋养细胞迁移能力的影响,深入了解缺氧环境诱发子痫前期发病的潜在分子机制。方法:体外贴壁培养的滋养细胞系(TEV-1),分别直接接种于24孔板进行细胞爬片和培养瓶内,分为常氧组和缺氧组。常氧组加新鲜DMEM/F12培养基,缺氧组加含终浓度为300μmol/L二氯化钴的培养基,然后在正常条件下(37℃,5%CO2)培养。两组细胞培养48 h后,分别收集细胞,采用Trizol法抽提mRNA,并进行RT-PCR,检测E-钙黏蛋白的mRNA表达水平;细胞爬片采用免疫组织化学法进行染色,检测E-钙黏蛋白的蛋白表达水平。统计数据分析两组差异性。结果:二氯化钴化学缺氧条件下培养后,滋养细胞E-钙黏蛋白的mRNA和蛋白表达水平均较常氧对照组升高,差异具有统计学意义(P<0.05)。结论:缺氧环境可能导致滋养细胞E-钙黏蛋白的mRNA和蛋白的表达水平升高,由此降低了滋养细胞的迁移能力。这可能是缺氧造成胎盘浅着床、继而发生子痫前期的重要分子机制。
OBJECTIVE: To investigate the expression of E-cadherin (E-cad) at the level of mRNA and protein in trophoblast cell line TEV-1 monolayer adherent cultured in hypoxia, Oxygen environment on the ability of trophoblast migration, in-depth understanding of the underlying molecular mechanism of hypoxic environment-induced preeclampsia. Methods: Trophoblast cells line (TEV-1) cultured in vitro were inoculated into 24-well plates for cell slide and culture flask respectively and divided into normoxia group and hypoxia group. The rats in normoxia group were treated with fresh DMEM / F12 medium, hypoxia group and medium supplemented with 300μmol / L cobalt chloride, and cultured under normal conditions (37 ℃, 5% CO2). After cultured for 48 hours, the cells were harvested and the mRNA was extracted by Trizol method. The mRNA expression of E-cadherin was detected by RT-PCR. The cell slides were stained with immunohistochemical method to detect E- Cadherin protein expression levels. Statistical analysis of two groups of differences. Results: The mRNA and protein expressions of E-cadherin in trophoblast cells were increased in the condition of hypochlorite chemical hypoxia compared with the normoxic control group (P <0.05). CONCLUSION: Hypoxia may result in increased expression of E-cadherin mRNA and protein in trophoblast cells, thereby reducing the ability of trophoblasts to migrate. This may be caused by hypoxia placentation shallow bed, followed by the important molecular mechanism of preeclampsia.