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

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、100 μmol/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，IC50）分别为308.4 μmol/L和322.0 μmol/L。10、100 μmol/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、100 μmol/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、100 μmol/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、100 μmol/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），100 μmol/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。

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

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 100 μmol/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 (IC50) values of forsythin for malignantly transformed esophageal epithelial cells and Eca-109 cells were 308.4 and 322.0 μmol/L, respectively. Forsythin at 10 and 100 μmol/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 100 μmol/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 100 μmol/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.