RNA干扰TAK1表达以增强三氧化二砷抑制Kasumi-1细胞增殖的作用及其机制研究
发布时间：2018-08-03 13:44
【摘要】：目的:探讨沉默转化生长因子β激活激酶(TAK1)基因表达对三氧化二砷(As_2O_3)抑制人t(8;21)急性髓系白血病细胞株Kasumi-1增殖的影响及其可能机制。方法:实验分为4组:对照组,TAK1特异siRNA转染Kasumi-1细胞组,As_2O_3作用组及两者联合处理的Kasumi-1细胞组。采用CCK-8法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,Western blot法检测TAK1、p-JNK及凋亡相关蛋白的表达。结果:在0.5-20μmol/L范围内,As_2O_3作用于Kasumi-1细胞24 h能够浓度依赖性地抑制Kasumi-1细胞增殖,24 h的IC_(50)值为(3.79±0.36)μmol/L;在0.5-10μmol/L范围内,As_2O_3作用于Kasumi-1细胞48 h能够浓度依赖性地抑制Kasumi-1细胞增殖;之后As_2O_3对Kasumi-1细胞的增殖抑制作用进入平台期,48 h的IC_(50)值为(2.38±0.17)μmol/L。TAK1siRNA转染组和As_2O_3 3.5μmol/L作用24 h组,Kasumi-1细胞的增殖抑制率分别为(10.86±1.64)%和(49.80±2.19)%,凋亡率分别为(8.47±0.75)%和(24.78±2.14)%,与对照组相比,差异有统计学意义(P0.05);两者联合作用于Kasumi-1细胞的增殖抑制率为(65.63±0.83)%,凋亡率为(68.97±2.94)%,与对照组和各单独组比较,差异均有统计学意义(P0.001)。TAK1siRNA转染和As_2O_3作用Kasumi-1细胞24 h能不同程度下调TAK1、p-JNK、c-Fos、c-Jun、BCL-2的表达,上调BAX和活化型(cleaved)Caspase-3、9的表达,与对照组比较差异均有统计学意义(P0.05),两者联合后该作用进一步加强(P0.05)。结论:利用RNA干扰技术沉默TAK1表达能够增强As_2O_3抑制Kasumi-1细胞增殖的作用,其原因可能是通过TAK1下游的JNK信号传导通路,以及线粒体途径诱导细胞凋亡实现的。
[Abstract]:Aim: to investigate the effect of silencing the expression of transforming growth factor 尾 -activated kinase (TAK1) on the inhibition of Kasumi-1 proliferation by arsenic trioxide (As_2O_3) and its possible mechanism. Methods: the experiment was divided into four groups: the control group was transfected with Kasumi-1 cells by TAK1 specific siRNA and the Kasumi-1 cells were treated with as _ 2O _ 3. CCK-8 assay was used to detect cell proliferation inhibition rate, and flow cytometry was used to detect apoptosis rate. Western blot assay was used to detect the expression of TAK1 p-JNK and apoptosis-related protein. Results: in the range of 0.5-20 渭 mol/L, the IC50 of Kasumi-1 cells was (3.79 卤0.36) 渭 mol / L in a dose-dependent manner, and the IC50 of Kasumi-1 cells was (3.79 卤0.36) 渭 mol / L in the range of 0.5-20 渭 mol/L, and the proliferation of Kasumi-1 cells was inhibited in a concentration-dependent manner in the range of 0.5-10 渭 mol/L. The IC50 of As_2O_3 on Kasumi-1 cells was (2.38 卤0.17) 渭 mol/L.TAK1siRNA transfection group and (10.86 卤1.64)% and (49.80 卤2.19)% of As_2O_3 3.5 渭 mol/L group respectively, and the apoptotic rate was (8.47 卤0.75)% and (24.78 卤2.14)% respectively. The inhibitory rate of proliferation and apoptosis were (65.63 卤0.83) and (68.97 卤2.94), respectively. Compared with the control group and the control group, the difference was statistically significant (P0.001) .TAK1 siRNA transfection and As_2O_3 could down-regulate the expression of TAK1p-JNKK-Fosc-Jun-BCL-2 in Kasumi-1 cells at 24 h. The up-regulated expression of BAX and activated (cleaved) Caspase-3 was significantly higher than that of the control group (P0.05), and the effect was further enhanced after the combination of the two groups (P0.05). Conclusion: silencing the expression of TAK1 by RNA interference technique can enhance the inhibitory effect of As_2O_3 on the proliferation of Kasumi-1 cells, which may be due to the JNK signal transduction pathway downstream of TAK1 and the mitochondrial pathway inducing apoptosis.
【作者单位】： 郑州大学附属肿瘤医院血液科河南省肿瘤医院血液科;郑州大学附属肿瘤医院中心实验室河南省肿瘤医院中心实验室;
【基金】：国家自然科学基金(81170520)
【分类号】：R733.71
本文编号：2161926
[Abstract]:Aim: to investigate the effect of silencing the expression of transforming growth factor 尾 -activated kinase (TAK1) on the inhibition of Kasumi-1 proliferation by arsenic trioxide (As_2O_3) and its possible mechanism. Methods: the experiment was divided into four groups: the control group was transfected with Kasumi-1 cells by TAK1 specific siRNA and the Kasumi-1 cells were treated with as _ 2O _ 3. CCK-8 assay was used to detect cell proliferation inhibition rate, and flow cytometry was used to detect apoptosis rate. Western blot assay was used to detect the expression of TAK1 p-JNK and apoptosis-related protein. Results: in the range of 0.5-20 渭 mol/L, the IC50 of Kasumi-1 cells was (3.79 卤0.36) 渭 mol / L in a dose-dependent manner, and the IC50 of Kasumi-1 cells was (3.79 卤0.36) 渭 mol / L in the range of 0.5-20 渭 mol/L, and the proliferation of Kasumi-1 cells was inhibited in a concentration-dependent manner in the range of 0.5-10 渭 mol/L. The IC50 of As_2O_3 on Kasumi-1 cells was (2.38 卤0.17) 渭 mol/L.TAK1siRNA transfection group and (10.86 卤1.64)% and (49.80 卤2.19)% of As_2O_3 3.5 渭 mol/L group respectively, and the apoptotic rate was (8.47 卤0.75)% and (24.78 卤2.14)% respectively. The inhibitory rate of proliferation and apoptosis were (65.63 卤0.83) and (68.97 卤2.94), respectively. Compared with the control group and the control group, the difference was statistically significant (P0.001) .TAK1 siRNA transfection and As_2O_3 could down-regulate the expression of TAK1p-JNKK-Fosc-Jun-BCL-2 in Kasumi-1 cells at 24 h. The up-regulated expression of BAX and activated (cleaved) Caspase-3 was significantly higher than that of the control group (P0.05), and the effect was further enhanced after the combination of the two groups (P0.05). Conclusion: silencing the expression of TAK1 by RNA interference technique can enhance the inhibitory effect of As_2O_3 on the proliferation of Kasumi-1 cells, which may be due to the JNK signal transduction pathway downstream of TAK1 and the mitochondrial pathway inducing apoptosis.
【作者单位】： 郑州大学附属肿瘤医院血液科河南省肿瘤医院血液科;郑州大学附属肿瘤医院中心实验室河南省肿瘤医院中心实验室;
【基金】：国家自然科学基金(81170520)
【分类号】：R733.71
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