Imaging of all immunohistochemical sections was performed Hedgehog antagonist using a Leica microscope electronic imager. The appearance of tan color or tan particles indicated a positive reaction in the cells. We performed IOD analysis on the sections in each group using Image Pro-plus v6.0 software to compare the differences between the group. 1.9 Statistical

analysis All data were analyzed using PASW 18.0 software and represented as . The variance analysis was adopted for comparisons between groups. P < 0.05 was considered to be statistically significant. Results 2.1 Effects of UTI and TAX on MDA-MB-231 cell proliferation Relative to the control group, the growth of MDA-MB-231 cells treated with UTI, TAX, and UTI+TAX for 24 h was significantly inhibited (P < 0.05; Table 1). The inhibitory effect increased in a time-dependent manner when the cells were treated for 48 learn more and 72 h (P < 0.01; Table 1). The strongest inhibitory effect was produced by co-treatment with both drugs

and the weakest effect occurred with UTI alone (UTI+TAX > TAX > UTI). The differences were statistically significant (P < 0.01; Table 1). Table 1 Effects of UTI and TAX on the proliferation of human breast cancer MDA-MB-231 cells in vitro (A570, )   24 h 48 h 72 h

Groups A value ( ) Inhibition rate (%) A value ( ) Inhibition rate (%) A value ( ) Inhibition rate (%) Control 1.086 ± 0.082 0 1.366 ± 0.042 0 1.881 ± 0.106 0 UTI 1.000 ± 0.067a 7.919 0.867 ± 0.102a 36.530 0.631 ± 0.067a 66.454 TAX 0.853 ± 0.051a,b 21.455 0.703 ± 0.043a,b 48.536 0.440 ± 0.063a,b 76.608 UTI+TAX 0.773 ± 0.041a,b,c 28.821 0.590 ± 0.059a,b,c 56.808 0.315 ± 0.068a,b,c 83.254 a P < 0.05 for all treatment groups versus control;b P < 0.01 for TXT and UTI+TAX groups versus UTI group;c P < 0.01 for UTI+TAX group versus Rolziracetam TAX group. 2.2 Effects of UTI and TAX on MDA-MB-231 cell apoptosis Compared to the control group (1.00), the level of apoptosis increased to 1.84 for the UTI group, 3.90 for the TAX group, and 6.79 for the UTI+TAX group (Table 2). Table 2 Apoptosis of MDA-MB-231 cells treated with different drugs Treatment Apoptotic rate(%) Fold increase Control 2.52 ± 0.53 0 UTI 7.16 ± 1.59 1.84 TAX 12.35 ± 1.88 3.90 UTI+TAX 19.64 ± 2.26 6.79 Data expressed as mean ± sd. Note: p < 0.05 among different treatments. 2.3 Expression of IL-6, IL-8, and TNF-α mRNA in MDA-MB-231 Treatment of MDA-MB-231 cells with both UTI and TAX down-regulated the expression of IL-6, IL-8, and TNF-α transcripts greater than treatment with either UTI or TAX alone (P < 0.05; Figure 1, Figure 2, Figure 3). Figure 1 Effects of UTI and TAX on IL-6 mRNA levels in MDA-MB-231 cells. Figure 2 Effects of UTI and TAX on IL-8 mRNA levels in MDA-MB-231 cells.