Shh和Gli-1在人乳腺癌MCF-7耐药细胞株中的表达及意义

目的 研究Shh和Gli-1在人乳腺癌耐药株中的表达情况，探讨Hedgehog信号通路与乳腺癌耐药的关系。方法 高浓度间歇诱导法建立人乳腺癌耐药细胞株MCF-7／PTX，MTT法检测紫杉醇（PTX）对MCF-7与MCF-7／PTX细胞的半数抑制浓度（IC50）。实时定量PCR（QPCR）检测MCF-7、MCF-7／PTX细胞中Shh、Gli-1 mRNA的表达。Western blotting检测MCF-7、MCF-7／PTX细胞中Shh、Gli-1蛋白的表达。结果 PTX 对 MCF-7细胞的IC50为（0.10±0.02）mg／L，对 MCF-7／PTX细胞的 IC50为（5.30±0.01）mg／L；耐药指数为53.0。Shh mRNA在MCF-7、MCF-7／PTX细胞中的表达量分别为0.78±0.12和1.45±0.56（P＜0.01）；Gli-1 mRNA在MCF-7、MCF-7／PTX细胞中的表达量分别为1.86±0.02和3.56±0.26（P＜0.01）。Shh 蛋白在MCF-7、MCF-7／PTX细胞中的表达量分别为0.58±0.06和1.03±0.22（P＜0.01）；Gli-1 蛋白在MCF-7、MCF-7／PTX细胞中的表达量分别为1.17±0.12和2.78±0.09（P＜0.01）。结论 人乳腺癌耐药细胞株MCF-7／PTX高表达Shh、Gli-1，化疗药物可能通过上调Hedgehog信号通路相关蛋白及基因介导乳腺癌耐药，针对该信号通路的靶向治疗将是克服乳腺癌耐药的一个新的选择。

Expression and significance of Shh and Gli-1 in drug-resistant cell strain of human breast cancer MCF-7

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 concen-tration( 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 chem-otherapy 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.