Genistein-induced apoptosis in breast cancer MDA-MB-231 cells

doi:10.3877/cma.j.issn.1674-0807.2013.05.003

Abstract

Abstract:

Objective

To explore the molecular mechanism of genistein-induced apoptosis in breast cancer MDA-MB-231 cells.

Methods

MDA-MB-231 cells were treated with 0, 5, 10, 20 μmol/L genistein for 24 h, respectively.CCK-8,Hoechst 33342 staining and flow cytometry were used to determine the effects of genistein on proliferation and apoptosis of MDA-MB-231 cells.The expressions of FADD, cleaved caspase-8,Fas, FasL in the protein level were measured by Western blotting.The expressions of Fas and FasL in the mRNA level were detected by real-time RT-PCR.The one-way ANOVA was conducted after homogeneity for variance was tested.

Results

The cell proliferation inhibition rate of MDA-MB-231 cells were (3.00±1.41)%,(14.02±1.57)%,(27.5±1.52)%,(48.90±1.44)% after the treatment of 0, 5, 10, 20 μmol/L genistein for 24 h respectively.Compared with 0 μmol/L group, the inhibition rate was increased in a concentration-dependent manner in the other 3 groups(F=528.119, P=0.000).The pairwise comparison showed the difference was statistically significant(all P＜0.05).The early apoptotic rate of MDA-MB-231 cells were (3.40±0.40)%,(9.34±1.34)%,(19.26±0.93)%,(27.41±1.12)% in each concentration group,respectively, after genistein treatment for 24 h.Compared with 0 μmol/L group, the early apoptotic rate was increased in a concentration-dependent manner in the other 3 groups(F=379.573,P=0.000).The pairwise comparison showed the difference was statistically significant(all P ＜0.05).Western blotting showed that genistein increased the protein expression levels of FADD, cleaved caspase-8 and FasL(F=368.621,456.744,419.129;all P=0.000), with no alteration in Fas protein level(F=0.800,P=0.528).Compared with 10 μmol/L group, FasL protein expression decreased in 20 μmol/L group(F=92.235,P=0.001).Real-time RT-PCR showed that genistein increased the mRNA expression of FasL(F= 646.983,P = 0.000), with alteration in Fas mRNA level(F = 1.556,P = 0.274).Compared with 10 μmol/L group, FasL mRNA expression also decreased in 20 μmol/L group(F=52.562,P=0.020).

Conclusion

Genistein induces apoptosis of breast cancer MDA-MB-231 cells by up-regulating FasL gene expression in Fas/FasL pathway.

Key words: Breast neoplasms, Apoptosis, Genistein, MDA-MB-231 cells, Fas/FasL pathway

Huan WANG, Chen-jia LU, Wei MA, Jun MAO, Pan-hong FAN, Wen-yue ZHAO, Lu WANG, Lian-hong LI. Genistein-induced apoptosis in breast cancer MDA-MB-231 cells[J]. Chinese Journal of Breast Disease(Electronic Edition), 2013, 07(05): 322-328.

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References 25

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