Effects of madecassic acid on proliferation,apoptosis,and autophagy of breast cancer MCF-7 cells
Objective To investigate the effects of madecassic acid(MA)on the proliferation,apoptosis,and autophagy of breast cancer MCF-7 cells.Methods MCF-7 cells were assigned into negative control group,MA(80,100,120,140,160 μmol/L)groups with different concentrations,and tamoxifen(10,20,30,40,50,35 μmol/L)groups with different concentrations,and they were treated for 24,36,and 48 h respectively.After the optimal concentration and time of MA anti-breast cancer effect were preliminarily determined,the cell viability was checked by MTT,and the corresponding half maximal inhibitory concentration(IC50)value was calculated.Flow cytometry was taken to detect apoptosis.JC-1 fluorescent probe was applied to check mitochondrial membrane potential changes.Western blot was used to determine the relative expression levels of proliferating cell nuclear antigen(PCNA),autophagy protein Beclin-1,microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ),and chelate 1(protein 62,p62).Transmission electron microscopy was used to detect autophagosomes.Results The optimal concentration of tamoxifen and MA were 35 μmol/L and 140 μmol/L respectively,and the optimal time was 48 h.Compared with the negative control group,the apoptosis rate,fluorescence relative intensity,and relative expression levels of LC3Ⅱ/Ⅰ and Beclin-1 protein in the MA 140 μmol/L group and tamoxifen 35 μmol/L group increased(P<0.05,P<0.01),and the relative expression levels of PCNA and p62 decreased(P<0.05,P<0.01).Compared with the MA 140 μmol/L group,the apoptosis rate,fluorescence relative intensity,and relative expression level of Beclin-1 protein in the tamoxifen 35 μmol/L group were significantly higher(P<0.01),and the relative expression level of PCNA protein was significantly lower(P<0.01).In the negative control group,the cell membrane of MCF-7 was intact,the nuclear membrane was clear,and the cell morphology was good;the cell membrane and nuclear morphology of the MA 140 μmol/L group were irregular,the mitochondrial matrix density was high,the ridge was expanded,and autophagosomes were visible;and the cell membrane of the tamoxifen 35 μmol/L group was partially dissolved and damaged,mitochondrial were swelling,matrix lysis was seen,and the double nucleoli and autophagosomes were visible.Conclusion MA can inhibit the proliferation of breast cancer MCF-7 cells and induce cell apoptosis and autophagy.
breast cancermadecassic acidproliferationapoptosisautophagyMCF-7 cells
万方数据
维普
