Objective To investigate the mechanism of Qizhen decoction on lipopolysaccharide (LPS) -induced epithelial mesenchymal transition (EMT) in colon cancer cells, and provide experimental evidence for its anti-cancer effects. Methods Human colon cancer Lo Vo cells were divided into 4 groups.The blank group was not treated. The model group, the experimental group and the control group were given 1 μg/ml LPS, 1 μg/ml LPS +800 μg/m L Qizhen decoction, and 1 μg/m L LPS + 100 μg/m L 5-fluorouracil (5-Fu) intervention for 48 h. The MTT assay was used to calculate the concentration of 50％ of tumor cell apoptosis induced by Qizhen decoction, which was IC50 (half maximal inhibitory concentration). The cell migration ability was observed by scratch test. The nuclear translocation of NF-κB p65 was detected by immunofluorescence. Western blot was used to detect the expression of E-cadherin and N-cadherin proteins. Results The IC50 value of Qizhen decoction induced apoptosis of tumor cells at 48 h was980.855 μg/m L. The scratch reduction areas of the blank group, the model group, the experimental group and the control group were (46.38 ± 5.15), (80.81 ± 3.18), (17.78 ± 9.87) and (16.95 ± 3.45), respectively. Compared with the model group, the scratch reduction areas of each group were extremely statistically significant (P ＜ 0.01). NF-κB p65 was more expressed than in cytoplasm in the blank group.In the model group, NF-κB p65 was observed to enter the nucleus, and nuclear translocation was obvious.NF-κB p65 was expressed both in the cytoplasm and nucleus in the control group. Compared with the blank group, the expression of E-cadherin in the experimental group (1.266 ± 0.065) and the control group (0.979 ± 0.183) were significantly higher than those in the model group (0.420 ± 0.071) (P ＜ 0.01).The expression of N-cadherin in the experimental group (0.518 ± 0.115) and the control group (1.275 ± 0.567) were significantly lower than those in the model group (2.287 ± 0.257) (P＜ 0.01).Conclusion Qizhen decoction can inhibit the occurrence of EMT in colon cancer cells by inhibiting the activation of NF-κB p65, thus achieving the effects of controlling tumor cell migration.%目的 研究芪贞汤对脂多糖(lipopolysaccharide,LPS)诱导结肠癌细胞上皮间充质转化(epithelial-mesenchymal transition,EMT)的机制,为其发挥抗癌作用提供实验依据.方法 将人结肠癌Lo Vo细胞分为4组,空白组不予干预,模型组、实验组和对照组分别予1μg/m L LPS、1μg/m L LPS+800μg/m L芪贞汤、1μg/m L LPS+100μg/m L 5-氟尿嘧啶(5-Fu)干预48小时.采用MTT实验计算芪贞汤诱导肿瘤细胞凋亡50％的浓度,即IC50(half maximal inhibitory concentration);划痕实验观察细胞迁移能力;免疫荧光法检测NF-κB p65核转位情况; Western blot检测E-钙黏蛋白(E-cadherin,E-cad)和N-钙黏蛋白(N-cadherin,N-cad)蛋白的表达.结果 芪贞汤在48小时诱导肿瘤细胞凋亡的IC50值为980.855μg/m L.空白组、模型组、实验组和对照组划痕缩小面积分别为(46.38±5.15)(80.81±3.18)(17.78±9.87)和(16.95±3.45);与模型组相比,各组划痕缩小面积皆有极显著统计差异(P ＜0.01).空白组NF-κB p65多于细胞浆中表达;模型组可以明显看到NF-κB p65入核,核转位明显;对照组NF-κB p65在包浆和细胞核中都有表达;实验组NF-κB p65在细胞核中表达相对较少,核转移情况下降.相对于空白组,实验组E-cadherin表达量(1.266±0.065)和对照组E-cadherin表达量(0.979±0.183)比模型组(0.420±0.071)显著升高,有极显著统计学差异(P ＜0.01);实验组N-cadherin表达量(0.518±0.115)和对照组N-cadherin表达量(1.275±0.567)比模型组(2.287±0.257)显著降低,有极显著统计学差异(P ＜0.01).结论 芪贞汤可通过抑制NF-κB p65活化而抑制结肠癌细胞EMT的发生,从而达到控制肿瘤细胞迁移的效果.
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