Notch1下调对三叶青黄酮抑制食管癌EC9706细胞迁移和侵袭的影响

Inhibitory Effect of Down-Regulation of Notch1 on Tetrastigma Hemsleyani Radix Flavone on Migration and Invasion of Esophageal Cancer EC9706 Cells

目的:观察三叶青黄酮(Tetrastigma Hemsleyani Radix flavone,THRF)对食管癌(esophagus cancer,EC)EC9706细胞增殖、迁移、侵袭能力及Notch1的影响,探讨其抗食管癌的可能作用机制。方法:体外培养EC9706细胞,以不同终质量浓度(0.5~20 g·L^(-1))三叶青黄酮进行处理,未加三叶青黄酮为空白组,以MW 167处理为MW167组。采用细胞计数试剂盒(cell counting kit-8,CCK-8),检测三叶青黄酮对EC9706细胞增殖的抑制作用,计算半数抑制浓度(50%inhibitory concentration,IC50);采用细胞黏附实验,划痕修复实验,transwell小室体外侵袭实验观察EC9706细胞黏附力、迁移、侵袭能力,同时采用免疫细胞化学和实时荧光定量PCR(Real-time PCR)检测Notch1蛋白和mRNA表达的变化。结果:与空白组比较,三叶青黄酮可明显升高EC9706细胞抑制率(P<0.05,P<0.01),三叶青黄酮可明显降低EC9706细胞黏附率、迁移率、侵袭细胞数,Notch1阳性细胞率和mRNA表达(P<0.05,P<0.01)。结论:三叶青黄酮能明显抑制EC9706细胞增殖,降低细胞迁移和侵袭能力,与其下调Notch1有关,可能是其抗食管癌的机制之一。

Objective： To investigate the effect of Tetrastigma Hemsleyani Radix flavone （THRF） on proliferation, migration, invasion and Notchl in esophageal cancer （EC） 9706 cells, and discuss its possible anti- esophageal cancer mechanism. Method： The EC9706 cells were cultured in vitro and treated with THRF with different concentrations （0.5 -20 g·L^-1） , with no THRF in blank group, and with MW 167 in positive control group. The 50% inhibitory concentration （ICs0） was measured by cell counting kit-8 （CCK-8）. The effect of THRF on adhesion rate was detected by the cell adhesion assay. The cell migration ability were evaluated by the wound healing assay. The cell invasion ability were evaluated by the transwell chamber assay. The expression of Notchl and Notch! mRNA expressions were detected by immunocytochemical analysis and Real-time PCR assay. Result ： THRF can significantly increase the inhibition rate of EC9706 cells, with statistically significant difference compared with blank group （P 〈 0.05, P 〈 0.01 ）. THRF can significantly inhibit the adhesion rate, the migration ability, the invasion cells, Notchl positive cell rate and Notchl mRNA expressions, with statistically significant difference compared with blank group （P 〈 0.05, P 〈 0.01 ）. Conclusion： THRF can inhibit the EC9706 cells viability, migration and invasion ability, its mechanism for anti-esophageal cancer may be correlated with down-regulation of Notchl.