桧木醇铁螯合物诱导乳腺癌细胞铁死亡机理研究

Mechanism of ferroptosis induced via hinokitiol iron chelate in breast cancer cells

为了探讨Fe(hino)3诱导乳腺癌细胞铁死亡机理,本文研究了Fe(hino)3对乳腺癌细胞的增殖活性、细胞内亚铁离子含量、丙二醛(malondialdehyde,MDA)含量、活性氧(reactive oxygen species,ROS)含量、细胞脂质过氧化变化和GSSG/GSH比值变化的影响;采用实时荧光定量PCR和Western blot研究了Fe(hino)3对乳腺癌细胞内Nrf2,HO-1,NQO1和Keap1基因和蛋白表达的影响,研究了纳米颗粒PLGA-Fe(hino)3对小鼠移植瘤增殖作用的影响.结果发现Fe(hino)3诱导乳腺癌细胞株MDA-MB-231和MCF7/ADR细胞铁死亡,铁死亡抑制剂DFO,Fer-1,Lip-1可显著抑制Fe(hino)3诱发的细胞死亡;细胞内亚铁离子含量、MDA含量、ROS含量、GSSG/GSH比值与Fe(hino)3的作用时间和浓度成正比;Fe(hino)3上调Nrf2,HO-1和NQO1基因和蛋白的表达,下调Keap1蛋白的表达;PLGA-Fe(hino)3抑制了小鼠移植瘤的增殖,PLGA-Fe(hino)3给药组移植瘤组织中Nrf2,HO-1蛋白表达量明显升高,移植瘤小鼠血清中MDA水平显著增加.以上结果表明,桧木醇铁螯合物诱导乳腺癌细胞铁死亡,抑制了乳腺癌细胞增殖.

To study Fe(hino)3-induced ferroptosis in breast cancer cells,the proliferative activity;levels of intracellular ferrous ion,malondialdehyde(MDA),and reactive oxygen species(ROS);and changes in cellular lipid peroxidation and GSSG/GSH ratio were examined.The effects of Fe(hino)3 on the expression of Nrf2,HO-1,NQO1,and Keap1 in breast cancer cells and the effect of PLGAFe(hino)3 nanoparticles on the proliferation of transplanted tumors in mice were investigated by performing real-time fluorescence quantitative polymerase chain reaction(q-PCR)and Western blot(WB).Fe(hino)3 induced ferroptosis in MDA-MB-231 and MCF7/ADR cells,whereas ferroptosis inhibitors,DFO,Fer-1,and Lip-1,significantly inhibited Fe(hino)3-induced cell death(P<0.05).Intracellular levels of ferrous ion,MDA,ROS,and GSSG/GSH were proportional to the time and concentration of Fe(hino)3.Fe(hino)3 upregulated the expression of Nrf2,HO-1,and NQO1 genes and proteins and downregulated the expression of the Keap1 protein(P<0.05).PLGA-Fe(hino)3 also decreased both the volume and weight of transplanted tumors in mice;further,the expression of Nrf2 and HO-1 proteins in transplanted tumor tissues was significantly increased in the PLGA-Fe(hino)3 administration group.MDA levels were significantly high in the serum of transplanted tumor mice.These results indicated that hinokiti iron chelate can induce ferroptosis and inhibit the proliferation of breast cancer cells.