| 刘小群,乔田奎,陈伟,袁素娟.含非甲基化二核苷酸的寡脱氧核苷酸7909对X射线诱导人肺腺癌A549细胞周期阻滞和凋亡的影响与ATM激酶的关系[J].中华放射医学与防护杂志,2012,32(3):270-273 |
| 含非甲基化二核苷酸的寡脱氧核苷酸7909对X射线诱导人肺腺癌A549细胞周期阻滞和凋亡的影响与ATM激酶的关系 |
| Role of ATM kinase in the effect of CpG ODN7909 on X-ray-induced G2/M phase arrest and apoptosis in A549 cells |
| 投稿时间：2011-11-20 |
| DOI：10.3760/cma.j.issn.0254-5098.2012.03.012 |
| 中文关键词: 含非甲基化二核苷酸的寡脱氧核苷酸 X射线 毛细血管扩张共济失凋突变激酶 细胞周期 凋亡 |
| 英文关键词:Unmethylated cytosine-phosphate-guanine oligodeoxynucleotides X-rays Ataxiatelangiectasia mutated kinase Cell cycle Apoptosis |
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| 目的 探讨含非甲基化二核苷酸的寡脱氧核苷酸(unmethylated cytosine-phosphate-guanine oligodeoxynucleotides, CpG ODN)7909对X射线诱导人肺腺癌A549细胞周期阻滞和凋亡的影响及毛细血管扩张共济失调突变(ATM)激酶磷酸化在这一过程中的作用。方法 人肺腺癌A549细胞用随机表法分为5组,对照组、CpG组、单纯照射组、CpG+X射线组和ATM siRNA+CpG+X射线组。采用集落形成法观察细胞存活率。流式细胞仪检测细胞周期和细胞凋亡。Western blot检测ATM激酶及其下游底物周期检测点激酶2(checkpoint kinase 2, Chk2)和P53蛋白的表达。结果 A549细胞经CpG ODN7909预处理后,在X射线照射下生长缓慢,集落形成明显减少,与单纯照射组比较,差异有统计学意义(t=13.41,P<0.05);增强了X射线诱导的人肺腺癌A549细胞G2/M期阻滞和凋亡,与单纯照射组比较,差异有统计学意义(t=17.32和7.71,P<0.05);明显增加了X射线诱导的ATM、Chk2及P53蛋白磷酸化水平;小分子干扰RNA暂时性抑制ATM的表达后,CpG ODN7909对X射线诱导G2/M期阻滞及凋亡的作用减弱,与CpG+X射线组比较,差异有统计学意义(t=26.84和2.98,P<0.05)。结论 CpG ODN7909在体外增强了X射线诱导的人肺腺癌A549细胞ATM激酶磷酸化,这一作用可能参与G2/M期阻滞和凋亡的调节。 |
| 英文摘要: |
| Objective To investigate the relationship between the effect of unmethylated cytosine-phosphate-guanine oligodeoxynucleotide(CpG ODN)7909 on X-ray-induced G2/M phase arrest and apoptosis and the phosphorylation of ATM kinase.Methods Human lung adenocarcinoma A549 cells were randomly classified into five groups, control, CpG group, irradiation group, CpG + irradiation group and ATM siRNA + CpG + irradiation group. Cell survival fraction was evaluated by clonogenic assay. Cell cycle and apoptosis were analyzed by flow cytometry. The expressions of ATM, checkpoint kinase-2 (Chk2) and p53 were detected with Western blot.Results Compared to the situation under X-ray irradiation alone, decreased cell clonogenic survival, prolonged G2/M arrest, and increased cell apoptosis were observed after the combination treatment with CpG ODN7909 and X-rays (t=13.41,17.32 and 7.71,P<0.05). Moreover, the phosphorylations on ATM, Chk2, and p53 were increased in the irradiated A549 cells that had been pre-treated with CpG ODN7909. After ATM siRNA interfering, abrogation of G2/M arrest, reduction of apoptosis, and decrease of Chk2 and p53 phosphorylation were found in A549 cells treated with CpG ODN7909 and X-rays (t=26.84,2.98,47.24 and 67.47,P<0.05). Conclusions CpG ODN7909 can enhance the X-ray-induced phosphorylation of ATM kinase in human lung adenocarcinoma cells in vitro, which might be involved in regulating G2/M phase arrest and apoptosis. |
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