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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。
Objective: To observe the effect of β-carotene on the proliferation, apoptosis, migration and cell cycle of esophageal squamous cell carcinoma EC1 cells. Methods: EC1 cells were treated with different concentrations (1,5,10,20,30 and 50μmol / L) of β-carotene for 12, 24, 36, 48, 60 and 72 h, respectively. The CCK- The optimal treatment time and concentration of β-carotene were screened. After that, EC1 cells were divided into 3 groups (negative control group, blank control group and experimental group), and Annexin V-FITC / PI double staining was used to detect cell apoptosis. Flow cytometry was used to detect cell cycle changes. Transwell migration assay was used to detect cell migration. Western blot was used to detect the expression of caveolin-1 (p-Akt) p-NF-κB, Bcl-2, Caspase-3, E-cadherin protein expression changes. Results: After screening, EC1 cells were treated with 50μmol / L β-carotene for 48 hours for subsequent experiments. Compared with the negative control group and the blank control group, the apoptosis rate of EC1 cells in experimental group was significantly increased (P <0.001), the proportion of cells in G0 / G1 phase increased (P <0.001), while the proportion of S phase cells decreased (P <0.001) (P <0.001), and the expression of E-cadherin was significantly increased (P <0.001). The expressions of Cav-1 and AKT signal pathways p-Akt, p-NF- -2 expression was significantly down-regulated, while Caspase-3 expression was activated (P <0.001). CONCLUSION: Beta-carotene can effectively promote the apoptosis of EC1 cells, suggesting that Cav-1-mediated AKT / NF-κB signaling pathway may play a role.