肿瘤坏死因子-α联合转化生长因子-β1作用于原代白血病细胞对Gli2表达的影响

Effects of TNF-α and TGF-β1 on Gli2 expression in primary leukemia cells

目的：证明肿瘤坏死因子-α（TNF-α）可以促进转化生长因子-β（TGF-β）降低原代白血病细胞Gli2的表达。方法 ：（1）Western blot检测白血病细胞Gli2蛋白的表达。（2）不同浓度TGF-β1和TNF-α分别作用于原代白血病细胞后,检测Gli2基因或蛋白以及TGF-βRⅠ和TGF-βRⅡ蛋白的表达。（3）TGF-β1、TNF-α及SIS3作用于原代白血病细胞24 h后,检测Gli2蛋白的表达。结果：（1）3株原代白血病细胞均有Gli2蛋白的表达。（2）1~10 ng/m L TGF-β1作用于原代白血病细胞12~24 h,与对照组相比Gli2基因m RNA表达降低明显;5 ng/m L TNF-α则没有显著变化。（3）5 ng/m L TNF-α作用于原代白血病细胞24 h后,与对照组相比TGF-βRⅠ和TGF-βRⅡ蛋白表达均有增高。（4）单独应用TGF-β以及联合应用TNF-α和TGF-β作用24 h后,与对照组相比Gli2基因的表达均降低明显,而且TNF-α＋TGF-β组Gli2基因的表达较TGF-β组降低更明显。结论：联合应用TNF-α和TGF-β比单独应用TGF-β能进一步减少原代白血病细胞Gli2基因的表达。

Objective To investigate whether TNF-α can promote, but TGF-β can reduce Gli2 expression in primary leukemia cells. Methods （1）Western blot was used to detect Gli2 protein expression in leukemic cells. （2）Primary leukemia cells were treated with TGF-β1 or TNF-α, and expressions of Gli2 and TGF-βR were detected. （3）Primary leukemia cells were treated with TGF-β1, TNF-α or/and SIS3, and Gli2 protein expression was detected. Results （1）The primary leukemia cells could express Gli2 protein. （2）Significant reduction of Gli2 mRNA was observed in 1 - 10 ng/mL TGF-β1-treated primary leukemia cells for 12 - 24 h, No significant differences of Gli2 mRNA was found between the control group and TNF-α group. （3）Protein expresson of TGF- βR I and TGF-βR 11 in 5 ng/mL TNF-α-treated primary leukemia cells for 24 h was higher than that of the control group. （4）Gli2 portein expression was lower in TGF-β1 with or without TNF-α-treated primary leukemia cells for 24 h than that in the control group, and Gli2 portein expression in the TGF-β plus TNF-α group was lower than that in TGF-β group. Conclusion Combinations of TGF-β and TNF-α can efficiently inhibit Gli2 expression than TGF-β alone in primary leukemia cells.