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目的探讨在白血病KG-1细胞株中转化生长因子-β(transforming growth factor-β,TGF-β)信号通路对Gli的调控作用。方法用0.1、1、10 ng/mL TGF-β1分别作用于KG-1细胞6、12和24 h,收集细胞,提取mRNA,检测Gli 2表达;5 ng/mL TGF-β1、5 ng/mL TGF-β1+5μmol/L SIS 3(specific inhibitor of Smad3)分别作用于KG-1细胞24 h,收集细胞,提取蛋白,检测Gli 2表达。结果 1、10 ng/mL TGF-β1分别作用于KG-1细胞6、12、24 h,其Gli 2的mRNA表达分别为0.78、0.51、0.16和0.73、0.59、0.15,与对照组比较,明显减少(P<0.05);与对照组比较,5 ng/mL TGF-β1组Gli 2蛋白表达明显降低,而5 ng/mL TGF-β1+5μmol/L SIS3组Gli 2蛋白表达明显升高。结论 TGF-β降低KG-1细胞Gli 2表达是通过TGF-β/Smad3途径介导的,不依赖于Ptch/Smo途径。
Objective To investigate the regulatory effect of transforming growth factor-β (TGF-β) signaling pathway on Gli in leukemia KG-1 cells. Methods KG-1 cells were treated with 0.1, 1, 10 ng / mL TGF-β1 for 6, 12 and 24 h, respectively. The cells were harvested and mRNA was extracted to detect the expression of Gli 2. 5 ng / mL TGF- TGF-β1 + 5μmol / L SIS 3 (specific inhibitor of Smad3) were respectively treated with KG-1 cells for 24 h. Cells were harvested and proteins were extracted for detection of Gli 2 expression. Results The mRNA expression of Gli 2 in KG-1 cells treated with 1, 10 ng / mL TGF-β1 for 6, 12 and 24 h were 0.78, 0.51, 0.16 and 0.73, 0.59 and 0.15 respectively. Compared with the control group, (P <0.05). Compared with the control group, the expression of Gli 2 protein in 5 ng / mL TGF-β1 group was significantly decreased, whereas the Gli 2 protein expression in 5 ng / mL TGF-β1 + 5 μmol / L SIS3 group was significantly increased. Conclusion TGF-βdown-regulates Gli 2 expression in KG-1 cells through the TGF-β / Smad3 pathway and does not depend on the Ptch / Smo pathway.