Silibinin affects transcription factors of cell cycle, apoptosis, angiogenesis and metastasis in breast cancer mice model: Targeting the hallmarks of cancer

Silibinin is a phyto-flavonoid compound and has been reported to exert anticancer effects on several tumors, including breast cancer. The present study exploited Flow cytometry, H&E staining, immunofluorescence (IF), and RT-PCR to extensively observe the effect of silibinin on the expression of cell-cycle and apoptosis-related transcription factors and genes in mice breast cancer cells 4T1. culture of 4T1 cell, MTT, flow cytometry, hematoxylin and eosin stains, immunofluorescent and Real Time PCR experiments were performed. silibinin caused a significant cytotoxicity, induction of apoptosis and cell cycle arrest in 4T1 cells, in vitro and in vivo. CD31 and HmgB1 expression showed a considerable decline. Evaluation of gene expression showed a marked decrease in Bcl-2, FoxC2, HoxB9, and FoxM1 genes, while a significant increase in TP53, P21, Bax, FoxF1, and HmgB1. Silibinin indicates the impact on inducing apoptosis by lower expression of Bcl-2, higher expression of Bax and TP53. While increased expression of P21 is thought to cause cell cycle arrest. In the meanwhile, under-expression of HoxB9, FoxC2, and FoxM1 and over-expression of FoxF1 caused restricted EMT, metastasis, angiogenesis and invasion in cancer.