TGF-β1体外诱导兔子宫内膜上皮细胞EMT模型

TGF-β1 induces EMT of rabbit endometrial epithelial cells in vitro

目的探讨转化生长因子-β1(TGF-β1)对兔子宫内膜上皮细胞的影响,以建立子宫内膜上皮细胞上皮间质转化(EMT)模型。方法分离培养新西兰大白兔子宫内膜上皮细胞,采用不同浓度(0、10、60、110μg/L)的TGF-β1诱导24 h和48 h,筛选TGF-β1最适诱导浓度和时间,建立EMT模型。观察细胞形态变化,分别采用CCK-8检测细胞的增殖抑制,Western blot及免疫组化检测波形蛋白(VIM)和E-钙黏蛋白(E-cadherin)表达变化。结果 60μg/L TGF-β1诱导细胞24 h能明显抑制兔子宫内膜上皮细胞的增殖作用,改变细胞形态,降低细胞密度使其向间质样形态发展,并下调E-cadherin蛋白和上调VIM蛋白的表达。结论 60μg/L TGF-β1诱导细胞24 h,能成功建立兔子宫内膜EMT模型。

Objective To investigate the effects of Transforming growth factor-β1(TGF-β1) on rabbit endometrial glandular epithelial cells to establish an epithelial-mesenchymal transition(EMT) model of endometrial epithelial cells. Methods Endometrial epithelial cells of New Zealand white rabbits were isolated and cultured, with different concentrations(0, 10, 60, 110 μg/L) of TGF-β1 induced 24 h and 48 h.EMT model was established by screening the optimal concentration and time of induction of TGF-β1.Morphological changes of cells were observed,CCK-8 was used to detect cell proliferation inhibition, and Western-blot and immunohistochemistry were used to detect changes in vimentin(VIM) and E-cadherin(E-cadherin) expression. Results 60 μg/L TGF-β1 inducing cell for 24 h significantly inhibited the proliferation of rabbit endometrial epithelial cells, changed cell morphology and decreased cell density to transform cells into mesenchymal morphology. Meanwhile, it down-regulated expression of E-cadherin protein and up-regulated expression of VIM protein. Conclusion The epithelial-mesenchymal transition model of rabbit endometrial epithelial cells can be successfully established with the concentration of 60 μg/L TGF-β1 inducing cells for 24 h.