β-胡萝卜素对食管鳞癌EC1细胞增殖、凋亡、迁移及细胞周期的影响

Effects of β-carotene on proliferation,apoptosis,migration and cell cycle of EC1 cells

目的:观察β-胡萝卜素对食管鳞癌EC1细胞增殖、凋亡、迁移及细胞周期的影响。方法:用不同浓度(1、5、10、20、30、50μmol/L)β-胡萝卜素分别处理EC1细胞12、24、36、48、60、72 h后,CCK-8法检测β-胡萝卜素对EC1细胞增殖率的影响,筛选β-胡萝卜素的最佳处理时间及浓度;随后,在此条件下将EC1细胞分为3组(阴性对照组、空白对照组和实验组),采用Annexin V-FITC/PI双染法检测细胞凋亡,流式细胞仪检测细胞周期的变化,Transwell迁移实验检测细胞迁移能力的变化,Western blot检测小窝蛋白1(Cav-1)、p-Akt、p-NF-κB、Bcl-2、Caspase-3、E-cadherin蛋白表达水平的变化。结果:经筛选,选用50μmol/Lβ-胡萝卜素作用EC1细胞48 h进行后续实验。与阴性对照组和空白对照组相比,实验组EC1细胞凋亡率显著提高(P<0.001),G0/G1期细胞比例增加(P<0.001),而S期细胞比例降低(P<0.001),发生迁移的细胞数量减少(P<0.001),且细胞黏附因子E-cadherin的表达显著增加(P<0.001),Cav-1及AKT信号通路关键蛋白p-Akt、p-NF-κB、Bcl-2表达量明显下调,而Caspase-3的表达被激活(P<0.001)。结论:β-胡萝卜素能够有效促进EC1细胞的凋亡,推测可能是通过抑制Cav-1介导的AKT/NF-κB信号通路发挥作用。

Aim： To investigate the effects of β-carotene on the proliferation,apoptosis,migration and cell cycle of human esophageal cancer EC1 cells and to study its related molecular mechanism. Methods： After treatment with different concentrations（ 1,5,10,20,30 and 50 μmol / L） of β-carotene for 12,24,36,48,60 and 72 h,the proliferation rate was tested by CCK-8 assay to determine the optimal time and concentration,EC1 cells were allocated into 3 groups： negative control group,blank control group and treatment group. Then the cell cycle and apoptosis were detected by FCM and Annexin V-FITC / PI staining respectively; the cell migration was measured by Transwell assay; effects of the β-carotene on protein expressions of Cav-1,p-Akt,p-NF-κB,Bcl-2,Caspase-3 and E-cadherin were detected by Western blot. Results： After screening,the optimal dose of β-carotene and treatment time for EC1 cells were determined as 50 μmol / L and48 h. After treatment with β-carotene,compared with negative control group and blank control group,EC1 cells in treatment group had a higher apoptosis rate（ P 0. 001）,the cell cycle changed significantly with an increase in G0/ G1 phase cells（ P 0. 001） and a decrease in S phase cells（ P 0. 001）,and the number of migrating cells was significantly less（ P 0. 001）. Meanwhile,the expression of E-cadherin in the treatment group was increased significantly（ P 0. 001）,p-Akt,p-NF-κB and Bcl-2 decreased,and Caspase-3 was activated（ P 0. 001）. Conclusion： β-carotene can effectively induce apoptosis of EC1 cells,which may be mediated by Cav-1 via AKT / NF-κB signaling pathway.