Abstract: Objective To investigate the expression of Shh，Gli-1 in human breast cancer resistant cell line and discuss the relationship between Hedgehog signaling pathway and drug resistance of breast cancer. Methods Drug-resistant strains of human breast cancer MCF-7 was established by high concentration intermittent induction. Paclitaxel（PTX）to half maximal inhibitory concentration（IC50）on MCF-7 and MCF-7／PTX cells was determined by MTT. QPCR was used to detect the mRNA expression of Shh，Gli-1 in MCF-7 and MCF-7／PTX cells. Protein expression of Shh，Gli-1 in MCF-7 and MCF-7／PTX cells was detected by Western blotting. Results IC50 of PTX for MCF-7 cells was（0.10+0.02）mg／L，and for MCF-7／PTX cell was（5.30+0.01） mg／L. The drug resistance index was 53.0. The expression of Shh mRNA in MCF-7 and MCF-7／PTX cells was 0.78 ± 0.12 and 1.45 ± 0.56（P＜0.01），and the expression levels of Gli-1 mRNA was 1.86±0.02 and 3.56±0.26（P＜0.01）. The expression level of Shh protein in MCF-7 and MCF-7／PTX cells was 0.58 ±0.06 and 1.03 ± 0.22（P＜0.01）. Gli-1 protein expression in MCF-7 and MCF-7／PTX cells was 1.17±0.12 and 2.78±0.09（P＜0.01）. Conclusion Shh，Gli-1 are highly expressed in MCF-7／PTX cells，indicating that chemotherapy may guide drug resistance of breast cancer by up-regulating the protein and gene of hedgehog signaling pathway. The target to hedgehog signaling pathway may be a new direction to overcome drug resistance.