Effect of Norathyriol on the Epithelial-mesenchymal Transition of HK-2 Cells Induced by TGF-β1
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摘要:
目的 研究芒果苷元对TGF-β1诱导的HK-2细胞EMT的影响。 方法 MTT法检测各浓度芒果苷元对HK-2细胞活力的影响；用TGF-β1诱导HK-2细胞成EMT模型，并给以芒果苷元干预，在TGF-β1诱导24 h时显微镜下拍照记录细胞形态变化，划痕实验法检测细胞迁移能力，Transwell法检测细胞侵袭能力，Western blot法检测纤连蛋白（fibronectin，FN）和Ⅰ型胶原（collagen type Ⅰ，Col Ⅰ）蛋白表达水平。 结果 （1）芒果苷元（10-9～10-5 mol/L）浓度下HK-2细胞活力均无明显变化（P > 0.05）；（2）与正常组相比，模型组细胞形态变长变梭，迁移和侵袭能力显著增强（ P < 0.05，0.01），FN和ColⅠ蛋白表达水平显著上调（ P < 0.05，0.01）；与模型组相比，芒果苷元组细胞能维持原来的鹅卵石形，迁移和侵袭能力显著下降（ P < 0.05，0.01），FN和ColⅠ蛋白表达水平显著下调（ P < 0.05）。 结论 芒果苷元能抑制TGF-β1诱导的HK-2细胞迁移和侵袭，阻止细胞形态的改变，下调细胞外基质成分FN和ColⅠ的蛋白表达，从而抑制EMT的发展。 Abstract:Objective To study the effects of norathyriol on epithelial-mesenchymal transition of HK-2 cells induced by TGF-β1. Methods MTT assay was used to detect the effect of norathyriol on HK-2 cell viability. The HK-2 cells were divided into normal group, model group, norathyriol （10-8, 10-7, 10-6 mol·L-1） groups and positive control （benzbromarone, 10-7 mol/L） group. The norathyriol and benzbromarone were incubated for 4 h before HK-2 cells were treated by TGF-β1 （10 ng/mL） to induce EMT. Then, the HK-2 cells were cultured for another 24 h or 48 h. Scratch assay was used to detect the migration motility of HK-2 cells. Transwell invasion assay was used to detect invasion ability of HK-2 cells. The proteins expression of fibronectin （FN） and collagen type Ⅰ （ColⅠ） were detected by western blot. Results （1） The norathyriol （10-9-10-5 mol/L）has no effects on the HK-2 cell viability, compared with the normal group （P > 0.05）. （2） Compared with the normal group, the cells morphology of the model group changed from the original cobblestone shape to a long fusiform shape, the migration rate and the invasive ability of the model group was significantly increased （ P < 0.01）. The protein expression of Col and FN in the model group was significantly increased. Compared with the model group, the cells morphology of norathyriol （10 -8, 10-7, 10-6 mol/L） and benzbromarone （10-7 mol/L） groups maintain thecobblestone shape partly, the migration rate （10-8, 10-7, 10-6 mol/L） and the invasive ability （10-6 mol/L） of norathyriol were significantly decreased （P < 0.01）; The protein expression of Col and FN were significantly down-regulated in the norathyriol （10 -6 mol/L） group and the benzbromarone group （P < 0.05, 0.01）. Conclusion Norathyriol can inhibit the migration and invasion of HK-2 cells induced by TGF-β1 and down-regulate the protein expression of fibronectin （FN） and collagen type Ⅰ （ColⅠ）. -
Key words:
- Norathyriol /
- TGF-β1 /
- HK-2 cell /
- Migration /
- Invasion /
- Epithelial-mesenchymal transition （EMT）
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图 1 芒果苷元对HK-2细胞活力的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；NL：芒果苷元；Ben：苯溴马隆。
Figure 1. Effect of norathyriol on HK-2 cell viability
图 2 芒果苷元对TGF-β1 诱导24 h后的HK-2细胞形态变化的影响（40×）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。
Figure 2. Effect of norathyriol on the morphological changes of HK-2 cells induced by TGF - β 1 for 24 h （40×）
图 3 芒果苷元对TGF-β1诱导后HK-2细胞迁移率的影响（40×）
Figure 3. Effect of norathyriol on the migration of HK-2 cells induced by TGF - β 1 （40×）
图 4 芒果苷元对TGF-β1诱导后的HK-2细胞的迁移率的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与模型组相比，*P < 0.05；**P < 0.01；与正常组相比，##P < 0.01。
Figure 4. Effect of norathyriol on the migration of HK-2 cells induced by TGF – β1 （ $\bar x \pm s$，n = 3）
图 5 芒果苷元对TGF-β1诱导的HK-2细胞侵袭能力的影响（100×）
Figure 5. Effect of norathyriol on invasion of HK-2 cells induced by TGF - β 1（100 ×）
图 6 芒果苷元对TGF-β1诱导的HK-2细胞侵袭能力的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与模型组相比，*P < 0.05；与正常组相比，##P < 0.01，one-way ANOVA。
Figure 6. Effect of norathyriol on invasion of HK-2 cells induced by TGF - β 1 （ $\bar x \pm s$，n = 3）
图 7 芒果苷元对TGF-β1诱导的HK-2细胞ColⅠ蛋白表达水平的影响（ $\bar x \pm s$，n = 3）
注：NOR：正常组；MOD：模型组；NL：芒果苷元；Ben：苯溴马隆。与正常组相比，#P < 0.05；与模型组比，*P < 0.05，**P < 0.01，one-way ANOVA。
Figure 7. Effect of norathyriol on expression of colⅠprotein in HK-2 cells induced by TGF-β1 （ $\bar x \pm s$，n = 3）
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