染料木黄酮在小细胞肺癌H446细胞中通过抑制FoxM1通路发挥抗肿瘤作用

Genistein exerts anti-tumor effects in small cell lung cancer H446 cells via suppressing the activity of FoxM1 pathw ay

目的研究染料木黄酮(genistein)在人小细胞肺癌H446细胞系中的抗肿瘤作用并探讨其分子机制。方法以不同浓度genistein(10、25、50、75、100μmol/L)处理H446细胞,采用CCK-8法、流式细胞术、质粒转染、Western blotting及Real-time PCR等技术检测genistein对细胞增殖、凋亡、细胞周期分布以及FoxM1及其下游靶基因表达的影响。结果 genistein引起H446细胞发生药物浓度相关性增殖抑制、凋亡增加及G2/M期细胞周期阻滞(P<0.05),此过程伴随FoxM1及其靶基因survivin、cyclinB、cdc25B的表达降低(P<0.05),而FoxM1过表达可部分拮抗genistein的抗肿瘤增殖作用(P<0.05)。结论 genistein可通过诱导细胞凋亡和细胞周期阻滞抑制H446细胞的增殖,而抑制FoxM1通路是其在小细胞肺癌中发挥抗肿瘤作用的机制之一。

Objective To investigate the influence of genistein on the growth of small cell lung cancer（SCLC） cell line H446 and the underlying molecular mechanisms. Methods Human SCLC cell line H446 were treated with various concentrations of genistein（10,25,50,75,100μmol/L）. CCK-8 assay, Flow cytometric analysis, Real-time PCR and Western blotting analysis were used to investigate the influences of genistein on cell growth, apoptosis, cell cycle progression and the mRNA and protein alterations of FoxM1 pathway. Results Genistein significantly inhibited the proliferation of H446 cells, accompanied by apoptosis and G2/M phase cell cycle arrest （P〈0.05）. Importantly, genistein led to attenuation on FoxM1 and decrease of its downstream genes, such as survivin, cyclin B, cdc25B, whereas p21 expression was increased （P〈0.05）. We also found that up-regulation of FoxM1 by cDNA transfection prior to genistein treatment reduced the inhibition of genistein-induced cell proliferation（P〈0.05）. Conclusion Genistein inhibits the proliferation of SCLC cells which is partly mediated through suppressing the activity of FoxM1 pathway.