鲁斯可皂苷元通过诱导铁死亡对胰腺癌细胞的抑制作用研究

Ruscogenin elements can inhibit pancreatic cancer cells by inducing iron death

目的 研究鲁斯可皂苷元(Ruscogenin, Rus)对胰腺癌的抑制作用,并初步研究其机制。方法 将SW1990和HPDE6-C7细胞进行接种,分别加入0.001,0.01,0.1,1,10和100μmol·L^(-1)浓度的Rus处理24,48和72 h后,用噻唑蓝(MTT)法测定细胞活力;用流式细胞术测定细胞死亡率;以普鲁士蓝染色光镜下测定细胞内亚铁含量;用DCFH-DA荧光探针法检测胞内活性氧(ROS)水平;以蛋白质印迹(Western Blot)法检测转铁蛋白(Transferrin)和铁调素(Ferroportin)表达。通过体内实验检测Rus对胰腺癌的抑制作用:SW1990细胞裸鼠成瘤后用Rus进行治疗,获得瘤体并称重并收集外周血和肾和肝组织进行毒性病理分析。结果 HPDE6-C7细胞中,0.1,1,10和100μmol·L^(-1)Rus处理72 h的细胞活性分别为(88.97±0.91)%,(85.65±1.55)%,(81.28±2.54)%和(65.20±5.93)%,SW1990细胞处理72 h的细胞活性分别为(83.79±1.26)%,(62.77±7.91)%,(36.73±7.45)%和(19.75±4.84)%,差异均有统计学意义(均P<0.05)。HPDE6-C7细胞中,0.1,1,10和100μmol·L^(-1)Rus处理48 h的细胞死亡率分别为(3.34±1.42)%,(10.68±2.19)%,(13.66±3.69)%和(21.05±6.53)%,SW1990细胞处理48 h的细胞死亡率分别为(8.95±3.43)%,(31.01±6.86)%,(63.23±7.37)%和(81.99±10.53)%,差异均有统计学意义(均P<0.05)。HPDE6-C7细胞中3和7μmol·L^(-1)浓度Rus处理24 h的细胞内亚铁水平分别为1.12±0.12和1.19±0.13,SW1990细胞处理24 h的细胞内亚铁水平分别为1.69±0.22和1.79±0.40,差异均有统计学意义(均P<0.05)。结论 Rus通过诱导铁死亡在胰腺癌细胞中发挥了抗癌功能。

Objective To explore the inhibitory effect of ruscogenin(Rus) on pancreatic cancer and its mechanism.Methods SW1990 and HPDE6-C7 cells were treated with Rus at concentrations of 0.001,0.01,0.1,1,10 and 100μmol·L^(-1) for 24,48 and 72 h,respectively,and cell viability was determined by MTT;cell mortality was determined by flow cytometry.The content of ferrous iron in cells was determined by Prussian blue staining.Intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescence probe.The expressions of Transferrin and Ferroportin were detected by Western Blot.The inhibitory effect of Rus on pancreatic cancer was examined by in vivo experiments:SW1990 cell nude mice were treated with Rus after tumor formation,tumors were obtained and weighed and peripheral blood and kidney and liver tissues were collected for toxic pathological analysis.Results In HPDE6-C7 cells,after 72 h treated with 0.1,1,10 and 100μmol·L^(-1) Rus,the cell activities were (88.97±0.91)%,(85.65±1.55)%,(81.28±2.54)%,(65.20±5.93)%,which in SW1990 cells were (83.79±1.26)%,(62.77±7.91)%,(36.73±7.45)%and (19.75±4.84)%,all with statistical significance (all P<0.05).In HPDE6-C7 cells,after 48 h treated with0.1,1,10 and 100μmol·L^(-1) Rus,the cell mortality were (3.34±1.42)%,(10.68±2.19)%,(13.66±3.69)%,(21.05±6.53)%,which in SW1990 cells were (8.95±3.43)%,(31.01±6.86)%,(63.23±7.37)%and (81.99±10.53)%,all with statistical significance (all P<0.05).HPDE6-C7 cells were treated with 3 and 7μmol·L^(-1) Rus for 24 h,the intracellular ferrous level were 1.12±0.12 and 1.19±0.13,SW1990 cells were 1.69±0.22 and 1.79±0.40,the difference was statistically significant (P<0.05).Conclusion Rus exerted anti-cancer function in pancreatic cancer cells by inducing ferroptosis.