| 周舟,张蕾,余晓东,邓朝晖,张广斌,余争平.γ射线诱导IEC-6细胞GSK-3β信号转导途径的激活[J].中华放射医学与防护杂志,2006,26(4):328-331 |
| γ射线诱导IEC-6细胞GSK-3β信号转导途径的激活 |
| Radiation-induced activation of GSK-3 β-coupled signal transduction pathway in IEC-6 cells |
| 投稿时间：2005-07-08 |
| DOI： |
| 中文关键词: γ射线 糖原合成酶激酶-3β caspase-9 caspase-3 |
| 英文关键词:γ-ray GSK-3 β caspase-9 caspase-3 |
| 基金项目:国家自然科学基金资助项目(30300072) |
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| 中文摘要: |
| 目的 观察60Coγ射线对IEC-6肠上皮细胞糖原合成酶激酶-3β(GSK-3β)信号转导途径的激活效应。方法 培养的IEC-6细胞接受6Gy60Coγ射线照射后,分别在照射后5、30、60和120min进行各项实验。应用RT-PCR法检测GSK-3β、Akt、caspase-9和caspase-3mRNA表达变化,Westernblot检测GSK-3β和Akt蛋白磷酸化的变化,分光法检测caspase-9酶活性。结果 6Gyγ射线照射后30min GSK-3βmRNA表达开始增高,60、120min GSK-3βmRNA均明显高于对照组;照射后30minGSK-3β蛋白磷酸化水平开始降低,60、120min GSK-3β蛋白磷酸化水平明显低于对照组水平;AktmRNA在γ射线照射后与对照组相比无明显改变;蛋白磷酸化水平在照射后30min开始降低,60和120min均明显低于对照组;caspase-9mRNA在照射后30min开始增高,60和120min均高于对照组,caspase-9酶活性变化趋势与mRNA一致,在照射后30min开始增高,并在辐射后60和120min逐渐升高;caspase-3mRNA在照射后60和120min高于对照组。结论 6Gyγ射线照射可以使GSK-3β上游的信号分子Akt磷酸化抑制,活性降低,进一步使GSK-3β在γ射线照射后磷酸化水平降低,被活化,从而激活GSK-3β信号转导途径,激活后的GSK-3β可激活其下游介导的与细胞凋亡相关的蛋白激酶家族的成员caspase-9、-3,从而造成IEC-6细胞的损伤。 |
| 英文摘要: |
| Objective To determine the effects of γ radiation on GSK-3 β-coupled signaling transduction pathway in IEC-6 cells. Methods The changes of GSK-3 β, Akt, caspase-9, and caspase-3 mRNA expressions in IEC-6 cells after 6 Gy γ-ray irradiation were determined using RT-PCR. GSK-3 β and Akt phosphorylation levels were determined by Western blot. Changes of caspase-9 enzymes activity were measured by colorimetric assay. Results GSK-3 β mRNA expressions at 30, 60 and 120 min after exposure were observed to be increased with 6 Gy γ-ray irradiation, while the corresponding levels of phosphorylated form of GSK-3 β decreased. Akt mRNA expression in the exposed group was not significantly different from that in the control group, but phosphorylation of Akt decreased at 30, 60, 120 min after exposure to 6 Gy γ-ray. Caspase-9 mRNA expression increased at 30, 60, 120 min and caspase-3 mRNA expression increased at 60, 120 min after the exposure. In consistence with the expression of caspase-9 mRNA, caspase-9 activity was markedly elevated at 30, 60, 120 min after exposure. Conclusion γ-ray irradiation induced significant activations of GSK-3 β signal transduction pathway and apoptosis-related caspase-9 and caspase-3 enzymes in IEC-6 cells. |
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