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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 observe the effect of Tetrastigma Hemsleyani Radix flavone (THRF) on proliferation, migration, invasion and Notch1 expression in esophagus cancer (EC) EC9706 cells and to explore its possible mechanism of action against esophageal cancer. Methods: EC9706 cells were cultured in vitro and treated with trichosanthin at different final concentrations (0.5-20 g · L -1). The untreated cells were treated with MW 167 for MW 167. The inhibitory effect of trichlorethylene on the proliferation of EC9706 cells was detected by cell counting kit-8 (CCK-8), and the 50% inhibitory concentration (IC50) was calculated. Cell adhesion assay, Repair experiment, transwell chamber invasion assay in vitro EC9706 cell adhesion, migration and invasion, while using immunocytochemistry and real-time PCR detection Notch1 protein and mRNA expression changes. Results: Compared with the blank group, cloverflower flavonoids could obviously increase the inhibitory rate of EC9706 cells (P <0.05, P <0.01), and clover leaf flavonoids could significantly reduce the adhesion rate, migration rate, number of invasive cells, Notch1 positive cells Cell rate and mRNA expression (P <0.05, P <0.01). CONCLUSION: Trilobin can significantly inhibit the proliferation of EC9706 cells and decrease the ability of cell migration and invasion, which may be related to its down-regulation of Notch1 and may be one of its mechanisms against esophageal cancer.