丹参酮ⅡA对胃癌细胞的抗癌作用及机制探讨

Study on the anticancer effect and mechanism of tanshinoneⅡA on gastric cancer cells

目的探讨丹参酮ⅡA(TⅡA)对胃癌细胞的抗癌作用及其机制。方法以胃癌细胞AGS、BGC-823为研究对象,基于MTT比色实验,计算TⅡA在胃癌细胞AGS、BGC-823中的半抑菌浓度(IC50)。选择合适浓度的TⅡA,采用流式细胞术分析TⅡA对细胞凋亡和死亡的影响。将胃癌细胞AGS、BGC-823分为对照组、TⅡA组和TⅡA+铁死亡抑制剂Fer-1组(TⅡA+Fer-1组),分别检测并比较每组细胞内谷胱甘肽(GSH)、半胱氨酸(Cys)、活性氧(ROS)及脂质过氧化水平。通过中药系统药理学和String数据库筛选并验证TⅡA潜在的作用靶点。采用蛋白质印迹法检测各组谷氨酸/胱氨酸转运蛋白(xCT)水平,采用实时荧光定量PCR检测TP53、溶质载体7家族11成员(SLC7A11)、前列腺素过氧化物内合酶2(PTGS2)的mRNA水平。结果TⅡA对胃癌细胞AGS、BGC-823具有良好的抗癌作用,IC50分别为2.880μg/mL和2.350μg/mL。TⅡA能抑制胃癌细胞AGS、BGC-823的生长,促进其凋亡和死亡。TⅡA能降低胃癌细胞AGS、BGC-823 GSH、Cys水平(P<0.05),升高ROS、脂质过氧化水平(P<0.05),最终引起细胞铁死亡。通过数据库分析发现,TP53是TⅡA重要的作用靶点。TⅡA能促进TP53的表达,抑制xCT的表达,Fer-1能削弱TⅡA对TP53、xCT表达的影响。加入TP53抑制剂后,TⅡA对SLC7A11、PTGS2、TP53的作用减弱(P<0.05)。结论TⅡA对胃癌细胞AGS、BGC-823具有良好的抗癌作用,可以通过TP53/xCT途径促进胃癌细胞铁死亡。

Objective To investigate the anticancer effect of tanshinoneⅡA(TⅡA)on gastric cancer cells and its mechanism.Methods Gastric cancer cells AGS and BGC-823 were used in this study,the semi inhibitory concentration(IC50)of TⅡA in gastric cancer cells AGS and BGC-823 were calculated based on MTT colorimetric assay.The appropriate concentration of TⅡA was selected.The effects of TⅡA on cell apoptosis and death were analyzed by flow cytometry.Gastric cancer cells AGS and BGC-823 were divided into control group,TⅡA group and TⅡA+ferroptosis inhibitor Fer-1 group(TⅡA+Fer-1 group).The levels of glutathione(GSH),cysteine(Cys),reactive oxygen species(ROS)and lipid peroxidation in each group were detected and compared.The potential targets of TⅡA were screened and verified by traditional Chinese medicine system pharmacology and String database.The levels of glutamate/cystine transporter(xCT)in each group were detected by Western blot,and the mRNA levels of TP53,solute carrier 7 family 11 members(SLC7A11),and prostaglandin peroxide endosynthase 2(PTGS2)were detected by real-time fluorescence quantitative PCR.Results TⅡA had a good anticancer effect on gastric cancer cells AGS and BGC-823 with IC50 of 2.880μg/mL and 2.350μg/mL,respectively.TⅡA could inhibit the growth and promote apoptosis and death of gastric cancer cells AGS and BGC-823.TⅡA treatment reduced GSH and Cys levels(P<0.05),increased ROS and lipid peroxidation levels(P<0.05),and finally induced ferroptosis in AGS and BGC-823 cells.Database analysis showed that TP53 was an important target of TⅡA.TⅡA promoted the expression of TP53 and inhibited the expression of xCT.Fer-1 attenuated the effects of TⅡA on the expression of TP53 and xCT.After adding TP53 inhibitor,the effects of TⅡA on SLC7A11,PTGS2,and TP53 were weakened(P<0.05).Conclusion TⅡA has a good anticancer effect on gastric cancer cells AGS and BGC-823,and it could promote ferroptosis of gastric cancer cells through TP53/xCT pathway.