【摘要】 目的探索白藜芦醇联合5-FU对宫颈癌细胞系HeLa细胞EMT的影响。方法 MTT实验检测白藜芦醇、5-FU以及联合使用对HeLa细胞增殖的影响;Transwell小室迁移实验检测各组细胞迁移能力;免疫荧光实验检测各组细胞中Vimentin蛋白表达变化;Western bloting实验检测各组细胞中N-cadherin、E-cadherin、Twist1、Snail1蛋白表达变化;Elisa实验检测各组细胞上清中MMP-2、MMP-9的表达。结果与单独使用白藜芦醇或5-FU相比,联合使用后HeLa细胞的IC50明显降低,提高了HeLa细胞对5-FU的敏感性。进一步发现与二者单独使用相比,联合使用导致HeLa细胞迁移能力显著下降。在分子水平上,白藜芦醇联合5-FU诱导细胞中E-cadherin蛋白表达更高,Vimentin、N-cadherin、Twist1、Snail1、MMP-2、MMP-9蛋白表达更低。结论与单独使用白藜芦醇或5-FU相比,白藜芦醇联合5-FU对HeLa细胞迁移能力的抑制增强,二者联合增强效应的分子机制在于联合使用对HeLa细胞EMT具有更强的逆转能力。更多还原

【Abstract】 Objective To investigate the effects of resveratrol combined with 5-FU on EMT transformation in cervical carcinoma cell line HeLa. Methods MTT assay was used to detect the effects of resveratrol, 5-FU and combination on the proliferation of HeLa cells. Transwell chamber migration assay was used to detect the migration ability of each group. Immunofluorescence assay was used to detect the expression of Vimentin protein in each group. The expressions of N-cadherin, E-cadherin, Twist1 and Snail1 in each group were detected by Western blotting(WB). The expressions of MMP-2 and MMP-9 in the supernatant of each group were detected by Elisa. Results Compared with resveratrol or 5-FU alone, the combined use of 5-FU significantly reduced the IC50 of HeLa cells and increased the sensitivity of HeLa cells to 5-FU. It was further found that the combined use resulted in a significant decrease in the migration ability of HeLa cells compared to the two alone. At the molecular level, resveratrol combined with 5-FU induced higher expression of E-cadherin protein, and Vimentin, N-cadherin, Twist1, Snail1, MMP-2, and MMP-9 proteins were lower. Conclusion The inhibition of HeLa cell migration ability is enhanced by resveratrol or 5-FU alone, and resveratrol combined with 5-FU. The molecular mechanism of the combined enhancement effect of the two is in combination with MDA-MB.-468 cell EMT transformation has a stronger reversal ability.更多还原