【摘要】 目的观察外源性转化生长因子β1(TGF-β1)对原代急性早幼粒细胞白血病细胞(APL)凋亡的影响,并探讨其可能机制。方法将APL细胞分为4组,对照组不加任何药物,实验组用终浓度为1、5、10 ng/mL TGF-β1处理(实验1、2、3组),分别处理24、48、72 h,采用瑞氏—吉姆萨染色法检测细胞凋亡,流式细胞仪检测细胞周期,免疫组化法检测TGF-β1、P27Kip1、Cyclin E、bcl-2蛋白,RT-PCR技术检测TGF-β1、P27Kip1、Cyclin E、bcl-2 mRNA。结果与对照组比较,实验1、2、3组24、48 h细胞凋亡率升高(P均<0.05);与同组24 h比较,实验1、2、3组48 h细胞凋亡率升高(P均<0.05)。对照组细胞培养48 h未见明显细胞周期阻滞;与对照组比较,实验1组48 h细胞周期阻滞于G1期,实验2、3组APL细胞周期明显阻滞于G1期。与对照组比较,实验2组TGF-β1、P27Kip1蛋白水平升高,Cyclin E、bcl-2蛋白降低(P均<0.05)。1’、2’β-actin条带亮度相同,实验2组(2道)P27Kip1mRNA条带亮度与对照组(1道)相同,P27Kip1mRNA水平较对照组变化不明显;1’、2’β-actin条带亮度相同,实验2组(2道)条带亮度明显高于对照组(1道);实验2组Cyclin E条带与对照组比较亮度减低,Cyclin E表达下调。结论外源性TGF-β1通过上调TGF-β1、P27Kip1及下调Cyclin E、bcl-2作用,诱导细胞发生凋亡,使细胞阻滞于G1期。更多还原

【Abstract】 Objective To observe the effect of exogenetic TGF-β1on apoptosis of primary acute promyelocytic leukaemia( APL) cells and to investigate the mechanism. Methods APL cells were divided into 4 groups: the control group,the experimental groups 1,2 and 3. The control group was not treated with any drugs,but the experimental groups 1,2 and 3 were separately treated with TGF-β1at the concentrations of 1,5,and 10 ng / mL for 24,48 and 72 h. The apoptosis was detected by Swiss-Jim staining method and cell cycle was detected by flow cytometry. The expression of TGF-β1,P27Kip1,Cyclin E and bcl-2 protein and mRNA in APL cells was detected by immunohistochemistry and RT-PCR. Results Compared with the control group,the apoptosis rate of the experimental groups 1,2 and 3 at 24 h and 48 h was increased( all P < 0. 05); in each group except the control group,the apoptosis rate at 48 h was higher than that at 24 h( all P < 0. 05). In the control group,after being cultured for 48 h,no cell cycle arrest was shown; compared with the control group,after being cultured for 48 h,the cell cycle of APL cells in the experimental group 1 was blocked in G1stage and the cell cycle of APL cells in the experimental groups 2 and 3 was significantly blocked in G1stage. Compared with the control group,the expression levels of TGF-β1and P27Kip1proteins were increased while Cyclin E,bcl-2 levels were decreased in APL cells( P < 0. 05). 1’, 2’ β-actin band had the same brightness,and the P27Kip1mRNA band was the same in the experimental group 2( 2’) and the control group( 1’),P27Kip1mRNA had no significant changes as compared with the control group; the brightness of Cyclin E band was reduced in the experimental group 2 as compared with that of the control group and the expression of Cyclin E was down-regulated. Conclusion Exogenetic TGF-β1can induce apoptosis and block the cell cycle in the G1stage by up-regulating the TGF-β1and P27Kip1,and down-regulating Cyclin E and bcl-2.更多还原