Effect of safflower polysaccharide combine cyclopamine on the proliferation and apoptosis of breast cancer cells

Abstract

Objective To investigate the effect of safflower polysaccharide combine cyclopamine on the proliferation and apoptosis of breast cancer cells. Methods Human breast cancer cell line MDA-MB-231 were cultured, cell proliferation and half maximal inhibitory concentration (IC50) calculation were detected after 0.05, 0.10, 0.20, 0.40, 0.80 mg/ml safflower polysaccharide and 2.5, 5.0, 10.0, 20.0, 40.0 μmol/L cyclopamine were added separately to cells for 24, 48, 72 h by cell counting kit-8 (CCK-8) test; the best concentration of safflower polysaccharide and cyclopamine were choiced according to the IC50; and the next test will be divided into control group, safflower polysaccharide group, cyclopamine group, safflower polysaccharide + cyclopamine group, cells proliferation in four groups were detected by CCK-8 test, apoptosis were detected by flow cytometry, and the expression of B cell lymphoma/leukemia-2 (bcl-2), bcl-2 associated X protein (bax), Fas, Gli protein was detected by Western blotting. Results IC50 calculation after safflower polysaccharide treated cells for 24, 48, 72 h cell were (0.89±0.04), (0.35±0.03), (0.14±0.02) mg/ml, IC50 calculation after safflower cyclopamine treated cells for 24, 48, 72 h cell were (56.30±1.23), (28.85±1.02), (15.26±0.87) μmol/L, survival rate after safflower polysaccharide and cyclopamine treated cells for 24, 48, 72 h were significantly lower than 0 hour According to the above concentration treatment, 0.8 mg/mL safflower polysaccharide and 20 μmol/L cyclopamine were used for a follow-up study; the cell survival rate in safflower polysaccharide group, cyclopamine group and safflower polysaccharide + cyclopamine group was 58.76%, 56.67%, 24.29%, the control group significantly higher than polysaccharide group (P=0.003), cyclopamine group (P=0.003) and safflower polysaccharide + cyclopamine group (P=0.000), the apoptosis rate in safflower polysaccharide group, cyclopamine group and safflower polysaccharide + cyclopamine group was 19.26%, 16.24%, 26.76%, the control group significantlyhigher than polysaccharide group (P=0.001), cyclopamine group (P=0.001) and safflower polysaccharide + cyclopamine group (P=0.000). cell survival rate in safflower polysaccharide+ cyclopamine group was significantly lower than safflower polysaccharide group (P=0.005)and cyclopamine group (P=0.005), the apoptosis rate in safflower polysaccharide+ cyclopamine group was significantly higher than safflower polysaccharide group (P=0.005) and cyclopamine group (P=0.004); the expression bcl-2 and Gli1 protein in safflower polysaccharide group, cyclopamine group, safflower polysaccharide+ cyclopamine group was significantly lower than the control group, the expression of bax and Fas protein was significantly higher than the control group; the expression of bcl-2, Gli1 protein in safflower polysaccharide+ cyclopamine group was significantly lower than the safflower polysaccharide group and the cyclopamine group, and bax and Fas protein expression was significantly higher than the safflower polysaccharide group and the cyclopamine group. Conclusion Safflower polysaccharide and Hedgehog pathway inhibitors were able to inhibit the proliferation of human breast cancer cell line MDA-MB-231, safflower polysaccharide and cyclopamine combination on cell proliferation and apoptosis were stronger than separate safflower polysaccharide and cyclopamine. Key words: Safflower polysaccharides; Hedgehog signal pathway; Cyclopamine; Breast cancer; Proliferation; Apoptosis