黄芩素与黄芩苷联合用药对口腔鳞癌细胞增殖、迁移的影响

Influence of baicalein combined with baicalin on the proliferation and migration of oral squamous cell carcinoma

目的探讨黄芩素与黄芩苷联合用药在体外条件下对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)CAL-27细胞的增殖、迁移能力的影响。方法应用MTT法检测黄芩素、黄芩苷及联合用药时对CAL-27细胞增殖能力的影响;应用划痕实验检测黄芩素、黄芩苷及联合用药时对CAL-27细胞迁移能力的影响;应用免疫荧光实验检测上皮间充质转化(epithelial-mesenchymal transition,EMT)的上皮标志物E钙粘蛋白(E-cadherin)与间充质标志物N钙粘蛋白(N-cadherin)的表达情况。结果MTT结果显示黄芩素与黄芩苷对CAL-27细胞的增殖均有抑制作用,随着浓度的增加,抑制率逐渐增强,而联合用药组对CAL-27细胞的抑制作用明显高于黄芩素及黄芩苷单药组(P=0.045,P=0.001),联合用药指数(Combination Index,CI)<1;划痕实验结果显示,24 h时,联合用药组和单药组的细胞愈合率均显著低于对照组(P=0.001,P=0.002,P=0.001),且联合用药组低于单药组(P=0.003,P=0.002);免疫荧光结果显示,黄芩素、黄芩苷及联合用药组与对照组相比,E-cadherin蛋白的表达升高,N-cadherin蛋白的表达降低,且联合用药组较单药组差异更为显著。结论黄芩素与黄芩苷均可抑制CAL-27细胞的增殖及迁移,且联合用药对CAL-27细胞增殖及迁移的抑制能力强于黄芩素与黄芩苷组,抑制作用可能与EMT的逆向表达有关。

Objective To investigate the effect of baicalein combined with baicalin on the proliferation and migration ability of oral squamous cell carcinoma(OSCC) CAL-27 cells in vitro. Methods The effects of baicalein, baicalin and their combination on the proliferation of CAL-27 cells were measured by MTT method. The effects of baicalin, baicalin and their combination on the migration of CAL-27 cells were tested by scratch test. Fluorescence experiments were performed to detect the expression of epithelial-mesenchymal transition(EMT) epithelial marker E-cadherin and mesenchymal marker N-cadherin. Results The MTT results showed that baicalein and baicalin could inhibit the proliferation of CAL-27 cells. With the increase of the concentration, the inhibition rate gradually increased, while the inhibitory effect of the combined drug group on CAL-27 cells was significantly higher than that of baicalein and baicalin monotherapy group(P=0.045, P=0.001), combined index(CI) <1. Scratch test results show that at 24 hours, the cell healing rates of the combined and monotherapy groups were significantly lower than the control group(P=0.001, P=0.002, P=0.001), and the combination drug group was lower than the single drug group(P=0.003, P=0.002). Immunofluorescence results showed that compared with the control group, in baicalein, baicalin and the combination drug groups, the expression of E-cadherin protein increased while the expression of N-cadherin protein decreased. And the difference in the combination group was more significant than the single drug group. Conclusion Both baicalin and baicalin can inhibit the proliferation and migration of CAL-27 cells, and the combined drug has a stronger ability to inhibit the proliferation and migration of CAL-27 cells than the baicalein and baicalin groups. The inhibition may be related to the reverse expression of EMT.