转化生长因子β1促进骨髓间充质干细胞迁移与上调Snail表达有关(英文)

Transforming growth factor beta 1 promotes the migration of mesenchymal stem cells via upregulation of Snail expression

背景:转化生长因子β1可促进骨髓间充质干细胞迁移及增殖,但机制尚不清楚。目的:观察转化生长因子β1对体外培养的骨髓间充质干细胞侵袭力的影响,以及对Snail、基质金属蛋白酶2表达的调节作用。方法:采用密度梯度离心结合贴壁法分离、培养大鼠骨髓间充质干细胞。用改良的Transwell小室检测0,0.5,1,2,5,10μg/L的转化生长因子β1对细胞迁移的影响。用脂质体转染针对Snail基因的特异性小分子干扰RNA至骨髓间充质干细胞,观察转染前后Snail和基质金属蛋白酶2的表达。结果与结论:外源性转化生长因子β1对骨髓间充质干细胞迁移的促进作用具有剂量依赖性,在2μg/L时达到最高。在此浓度下,Snail、基质金属蛋白酶2mRNA表达明显增高。Snail基因沉默可以明显抑制转化生长因子β1对基质金属蛋白酶2表达的促进作用。提示转化生长因子β1可通过促进骨髓间充质干细胞基质金属蛋白酶2的表达提高其迁移能力,此作用与上调Snail表达有关。

BACKGROUND： Transformation growth factor β1 （TGF-β1） can promote bone marrow mesenchymal stem cells （BMSCs） migration and proliferation, but the underlying mechanisms remain unclear.OBJECTIVE： To observe the invasiveness of TGF-β1 on BMSCs cultured in vitro, and to investigate regulatory effect on Snail and matrix metalloproteinase 2 （MMP-2） expression.METHODS： Rat BMSCs were isolated and cultured with density gradient centrifugalization and adherence method. The influence of different concentrations of TGF-β1 on the BMSC migration was detected using the modified Transwell chambers. Small interfering RNA for Snail gene was synthesized and transfected into BMSCs by liposomel before TGF-β1 was treated, and the expression of Snail and MMP-2 before and after transfection were measured by western blot assay.RESULTS AND CONCLUSION： The exogenous TGF-β1 can induce a dose-dependent increase in cell migration, which peaked at 2 μg/L. The expression levels of Snail mRNA and MMP-2 mRNA were significantly increased after 2 μg/L TGF-β1 treatment. Snail gene can effectively inhibit the expression of MMP-2 promoted by TGF-β1. Experimental findings indicate that TGF-β1 could increase the MMP-2 expression and then promote the BMSCs migration through the upregulation of the Snail expression.