连翘苷对食管上皮恶性转变细胞和食管癌Eca-109细胞的抑制作用及机制  

Inhibitory Effect and Mechanism of Forsythin on Malignantly Transformed Esophageal Epithelial Cells and Esophageal Cancer Eca-109 Cells

作　　者：郭子琪 王少康[1] 桂兰兰 夏惠[1] 孙桂菊[1] GUO Ziqi;WANG Shaokang;GUI Lanlan;XIA Hui;SUN Guiju(School of Public Health,Southeast University,Nanjing 210009,China)

机构地区：[1]东南大学公共卫生学院,江苏南京210009

出　　处：《食品科学》2022年第15期176-184,共9页Food Science

摘　　要：目的:探究连翘苷对食管上皮恶性转变细胞和食管癌Eca-109细胞增殖、周期、凋亡、迁移、侵袭的影响,分析其对这两种细胞的抑制效果及其作用机制。方法:取食管上皮恶性转变细胞和食管癌Eca-109细胞,使用不同浓度(10、100μmol/L)的连翘苷处理干预细胞24 h,采用细胞计数试剂盒-8(cell counting kit-8,CCK8)法检测细胞增殖,流式细胞术检测细胞周期、凋亡,Transwell实验检测细胞迁移、侵袭,蛋白免疫印迹法(Western blotting)检测细胞中磷脂酰肌醇-3-激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,PKB),(也称为AKT)、磷酸化磷脂酰肌醇3-激酶(phosphorylated phosphatidylinositol-3-kinase,p-PI3K)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-PKB)(也称为p-AKT)、p21蛋白、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和基质金属蛋白酶9(matrix metallopeptidase 9,MMP9)表达水平。结果:连翘苷对食管上皮恶性转变细胞和食管癌Eca-109细胞的半数抑制浓度(50%inhibitory concentration,IC_(50))分别为308.4μmol/L和322.0μmol/L。10、100μmol/L连翘苷可显著抑制食管上皮恶性转变细胞和Eca-109细胞增殖(P<0.05),对人食管正常上皮细胞无毒性作用。10、100μmol/L连翘苷可诱导食管上皮恶性转变细胞和Eca-109细胞周期改变,促进细胞凋亡。10、100μmol/L连翘苷可显著抑制食管上皮恶性转变细胞和Eca-109细胞迁移及侵袭(P<0.05)。10、100μmol/L连翘苷可显著降低食管上皮恶性转变细胞和Eca-109细胞中p-PI3K、p-AKT、Bcl-2和MMP9蛋白的表达水平(P<0.05),100μmol/L连翘苷可显著提高食管上皮恶性转变细胞和Eca-109细胞中p21蛋白的表达水平(P<0.05)。结论:连翘苷可显著抑制食管上皮恶性转变细胞和食管癌Eca-109细胞增殖、迁移、侵袭,其机制可能与PI3K/AKT通路失活有关。Objective:The inhibitory effect of forsythin on the proliferation,cell cycle,apoptosis,migration and invasion of malignantly transformed esophageal epithelial cells and esophageal cancer Eca-109 cells was explored,and the underlying mechanism was analyzed.Methods:Malignantly transformed esophageal epithelial cells and Eca-109 cells were treated with 10 and 100μmol/L forsythin for 24 h.Cell proliferation was detected by cell counting kit-8(CCK-8).Cell cycle and apoptosis were detected by flow cytometry.Cell migration and invasion were detected by Transwell method.The expression levels of phosphoinositide 3-kinase(PI3K),protein kinase B(PKB,also known as AKT),phosphorylated phosphatidylinositol-3-kinase(p-PI3K),phosphorylated protein kinase B(p-PKB,also knoun as p-AKT),p21,B-cell lymphoma-2(Bcl-2),and matrix metallopeptidase 9(MMP9)in cells were detected by Western blotting.Results:The 50%inhibitory concentration(IC_(50))values of forsythin for malignantly transformed esophageal epithelial cells and Eca-109 cells were 308.4 and 322.0μmol/L,respectively.Forsythin at 10 and 100μmol/L could significantly inhibit the proliferation of the two cell lines(P<0.05),but was nontoxic to normal esophageal epithelial cells.Forsythin at 10 and 100μmol/L could alter cell cycle,promote cell apoptosis,significantly inhibit cell migration and invasion(P<0.05),and reduce the expression levels of p-PI3K,p-AKT,Bcl-2 and MMP9 proteins(P<0.05).Forsythin at 100μmol/L could significantly increase the expression level of p21 protein in both cell lines(P<0.05).Conclusion:Forsythin can significantly inhibit the proliferation,migration and invasion of esophageal epithelial malignant transformed cells and Eca-109 cells,and the mechanism may be related to the inactivation of the PI3K/AKT pathway.

关 键 词：连翘苷 食管上皮恶性转变细胞 ECA109细胞 PI3K/AKT 

分 类 号：R151.41[医药卫生—营养与食品卫生学]