电离辐射对胃肠道间质瘤细胞增殖以及Ano1蛋白表达的影响

Effects of Ionizing Radiation on the Proliferation of GIST Cells and the Expression of Ano1 Protein

目的探究电离辐射后胃肠道间质瘤(gastrointestinal stromaltumor,GIST)细胞增殖活性和凋亡以及电离辐射后GIST中Ano1分子的mRNA和蛋白表达。方法实验细胞为人胃肠道间质瘤细胞株(GIST-T1),实验分组为未照射组、2 Gy照射组、5 Gy照射组、7 Gy照射组,观察时间点分别为照射后1 d、2 d、3 d、4 d、5 d。通过CCK8法和流式细胞仪检测电离辐射后GIST细胞增殖活性和凋亡,以RT-PCR和Western Blot方法检测电离辐射后GIST中Ano1分子的mRNA和蛋白表达。结果GIST细胞在接受不同剂量的电离辐射急性照射后,生长受到抑制,且有凋亡表现,Ano1蛋白的表达出现了先下降、后回升的变化。随着照射剂量增加,Ano1表达下降的程度越大、到达谷值时间越短,且Ano1的表达恢复速度越慢。结论GIST细胞增殖活性的抑制程度和细胞凋亡率与照射剂量和照射后时间呈依赖关系,Ano1蛋白的表达出现了先下降、后回升的变化。提示Ano1可能是GIST的放疗靶点。

Objective To explore the effect of ionizing radiation on the proliferation activity and apoptosis of gastroin⁃testinal stromal tumor(GIST)cells as well as the expressions of mRNA and protein of Ano1 molecules in GIST cells.Methods GIST-T1 cells were applied in experiment and were divided,based on different radiation dose,into 4 groups:non-radiation group,2 Gy group,5 Gy group,and 7 Gy group;the observation time points were 1d,2d,3d,4d and 5d af⁃ter radiation;the proliferation activity and apoptosis of GIST cells after ionizing radiation were detected by CCK8 meth⁃od and flow cytometry and the mRNA and protein expressions of Ano1 molecules in GIST cells after ionizing radiation were detected by RT-PCR method and Western Blot method.Results After acute exposure to ionizing radiation,the in⁃hibition of growth and the apoptosis were found in GIST cells;the expression of Ano1 protein had first decreased and then recovered after acute exposure to ionizing radiation;radiation dose was positively correlated with the degree of Ano1 expression decline and negatively correlated with the time to reach the trough and the recovery rate of Ano1 ex⁃pression.Conclusions The inhibition of cell proliferation activity and the rate of apoptosis in GIST cells are dependent on the dose and duration of radiation;the decline and then recovery of Ano1 protein indicates that Ano1 is a possible ra⁃diotherapy target of GIST.