發(fā)布時(shí)間：2018-08-03 13:44

【摘要】：目的:探討沉默轉(zhuǎn)化生長(zhǎng)因子β激活激酶(TAK1)基因表達(dá)對(duì)三氧化二砷(As_2O_3)抑制人t(8;21)急性髓系白血病細(xì)胞株Kasumi-1增殖的影響及其可能機(jī)制。方法:實(shí)驗(yàn)分為4組:對(duì)照組,TAK1特異siRNA轉(zhuǎn)染Kasumi-1細(xì)胞組,As_2O_3作用組及兩者聯(lián)合處理的Kasumi-1細(xì)胞組。采用CCK-8法檢測(cè)細(xì)胞增殖抑制率,流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡率,Western blot法檢測(cè)TAK1、p-JNK及凋亡相關(guān)蛋白的表達(dá)。結(jié)果:在0.5-20μmol/L范圍內(nèi),As_2O_3作用于Kasumi-1細(xì)胞24 h能夠濃度依賴性地抑制Kasumi-1細(xì)胞增殖,24 h的IC_(50)值為(3.79±0.36)μmol/L;在0.5-10μmol/L范圍內(nèi),As_2O_3作用于Kasumi-1細(xì)胞48 h能夠濃度依賴性地抑制Kasumi-1細(xì)胞增殖;之后As_2O_3對(duì)Kasumi-1細(xì)胞的增殖抑制作用進(jìn)入平臺(tái)期,48 h的IC_(50)值為(2.38±0.17)μmol/L。TAK1siRNA轉(zhuǎn)染組和As_2O_3 3.5μmol/L作用24 h組,Kasumi-1細(xì)胞的增殖抑制率分別為(10.86±1.64)%和(49.80±2.19)%,凋亡率分別為(8.47±0.75)%和(24.78±2.14)%,與對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(P0.05);兩者聯(lián)合作用于Kasumi-1細(xì)胞的增殖抑制率為(65.63±0.83)%,凋亡率為(68.97±2.94)%,與對(duì)照組和各單獨(dú)組比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.001)。TAK1siRNA轉(zhuǎn)染和As_2O_3作用Kasumi-1細(xì)胞24 h能不同程度下調(diào)TAK1、p-JNK、c-Fos、c-Jun、BCL-2的表達(dá),上調(diào)BAX和活化型(cleaved)Caspase-3、9的表達(dá),與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05),兩者聯(lián)合后該作用進(jìn)一步加強(qiáng)(P0.05)。結(jié)論:利用RNA干擾技術(shù)沉默TAK1表達(dá)能夠增強(qiáng)As_2O_3抑制Kasumi-1細(xì)胞增殖的作用,其原因可能是通過TAK1下游的JNK信號(hào)傳導(dǎo)通路,以及線粒體途徑誘導(dǎo)細(xì)胞凋亡實(shí)現(xiàn)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 鹵0.36) 渭 mol / L in a dose-dependent manner, and the IC50 of Kasumi-1 cells was (3.79 鹵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 鹵0.17) 渭 mol/L.TAK1siRNA transfection group and (10.86 鹵1.64)% and (49.80 鹵2.19)% of As_2O_3 3.5 渭 mol/L group respectively, and the apoptotic rate was (8.47 鹵0.75)% and (24.78 鹵2.14)% respectively. The inhibitory rate of proliferation and apoptosis were (65.63 鹵0.83) and (68.97 鹵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.
【作者單位】： 鄭州大學(xué)附屬腫瘤醫(yī)院血液科河南省腫瘤醫(yī)院血液科;鄭州大學(xué)附屬腫瘤醫(yī)院中心實(shí)驗(yàn)室河南省腫瘤醫(yī)院中心實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(81170520)
【分類號(hào)】：R733.71

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