紫草素对胃癌细胞糖酵解代谢及生物学特性的影响

Effects of Shikonin on Glycolytic Metabolism and Biological Characteristics of Gastric Cancer Cells

目的:研究紫草素对胃癌细胞糖酵解代谢及生物学特性的影响。方法:经不同浓度的紫草素处理后,CCK-8法检测正常胃上皮GES-1细胞及胃癌SGC-7901细胞增殖;划痕实验和Transwell实验检测SGC-7901细胞迁移和侵袭能力,采用流式细胞仪分析SGC-7901细胞凋亡;采用相应试剂盒测定GES-1和SGC-7901细胞乳酸、丙酮酸和柠檬酸水平;Western Blotting检测SGC-7901细胞HK2、LDHA、c-Myc蛋白表达。CCK-8法和Western Blotting检测外源性乳酸作用下紫草素对SGC-7901细胞活力和c-Myc蛋白表达的影响。结果:紫草素能显著抑制SGC-7901细胞增殖、迁移、侵袭,并诱导其凋亡,降低SGC-7901细胞内乳酸和丙酮酸水平,显著升高SGC-7901细胞内柠檬酸水平。Western Blotting结果显示,紫草素处理后SGC-7901细胞HK2、LDHA、c-Myc蛋白表达显著降低,且其作用呈浓度依赖性。紫草素对GES-1细胞活力及乳酸、丙酮酸水平无明显影响。加入外源性乳酸后,紫草素作用下的SGC-7901细胞活力升高,c-Myc蛋白表达上调。结论:紫草素可抑制胃癌SGC-7901细胞增殖、迁移、侵袭并诱导其凋亡,其机制与抑制c-Myc信号轴及改变糖酵解代谢密切相关。

Objective:To study the effect of shikonin on glycolysis metabolism and biological characteristics of gastric cancer cells.Methods:The proliferation of normal gastric epithelial GES-1 cells and gastric cancer SGC-7901 cells was detected by CCK-8 method after treatment with different concentrations of shikonin.The migration and invasion ability of SGC-7901 cells were detected by scratch assay and Transwell assay.The apoptosis of SGC-7901 cells was analyzed by flow cytometry.The levels of lactic acid,pyruvate and citric acid in GES-1 cells and SGC-7901 cells were determined by the corresponding kit.The protein expressions of HK2,LDHA and c-Myc in SGC-7901 cells were detected by Western Blotting.CCK-8 method and Western Blotting CCK-8 method were used to detect the effects of shikonin on the cell viability and c-Myc protein expression of SGC-7901 under exogenous lactic acid.Results:Shikonin significantly inhibited the proliferation,migration and invasion of SGC-7901 cells,induced cell apoptosis,decreased the levels of lactic acid and pyruvate in SGC-7901 cells,and significantly increased the levels of citric acid in SGC-7901 cells.Western Blotting results showed that the protein expressions of HK2,LDHA and c-Myc in SGC-7901 cells were significantly decreased after treatment with shikonin in a concentration-dependent manner.The activity of GES-1 cells and the levels of lactic acid and pyruvate in normal gastric epithelium were not affected by schillin.After the addition of exogenous lactic acid,the activity of SGC-7901 cells was increased and the protein expression of c-Myc was up-regulated.Conclusion:Shikonin can inhibit proliferation,migration,invasion and induce apoptosis of SGC-7901 cells,and its mechanism is closely related to inhibiting c-Myc signal axis and altering glycolytic metabolism.