关键词: 异嗪皮啶, 乳腺癌干细胞, WST-8, 细胞凋亡, Bcl-2, Caspase

Abstract:

AIM: To investigate the effects of isofraxidin from sarcandra glabra on the expression levels of Bcl-2, Caspase-3 and Caspase-8 which are related with the apoptosis in breast cancer stem cells.  METHODS: The breast cancer stem-like cells were enriched from MDA-MB-231 cell lines using the serum-free culture method, and the CD44⁺/CD24 ^-/low subpopulation was sorted using the flow cytometry. After 0, 17, 50, 150 and 450 μmol/L isofraxidin were administrated to each group, the cell viability were detected using CCK-8 reagent, the width of cell wound was continuously recorded via microscopic photograph, and the total cell apoptosis rates were recorded using Annexin V-FITC/PI staining. The mRNA and protein levels of Bcl-2, Caspase-3 and Caspase-8 were measured using Real-time quantitative PCR and Western blot, respectively. RESULTS: Significantly lower rates of proliferation were observed in 50, 150, and 450 μmol/L isofraxidin group at 24, 48 and 72 h. 50, 150 and 450 μmol/L isofraxidin given for 48 h results in the decline of cell migration and increase in the rate of apoptotic. For the levels of mRNA and protein, Bcl-2 gene of 50, 150 and 450 μmol/L group was higher than that of the control, nevertheless, caspase-3 and caspase-8 gene of these groups were lower than those of the control. All the data showed a clear-cut dose-effect relationship. CONCLUSION: The up-regulation of Bcl-2 and the down-regulation of Casepase-3/8 genes are related with the apoptosis, proliferation inhibition and reduced migration of breast cancer stem cells after isofraxidin treatment.

Key words: isofraxidin, tumor stem cells, WST-8, apoptosis, Bcl-2, Caspase