DYRK1B基因对食管鳞癌细胞放射敏感性影响的研究

Effect of DYRK1B Gene on Radiosensitivity of Esophageal Squamous Cell Carcinoma ECA109 Cells

[目的]探讨DYRK1B对食管鳞癌放射敏感性的影响及潜在机制,为提高食管鳞癌患者放射敏感性提供理论依据。[方法]用脂质体Lipofectamine 3000分别包裹siNC、siDYRK1B转染食管鳞癌细胞ECA109,48小时后收集细胞RT-PCR和蛋白印迹实验检测ECA109细胞中DYRK1B表达水平。同时取转染siNC、siDYRK1B的ECA109细胞给予放射处理,用克隆形成实验检测经不同剂量放射处理后的细胞存活情况并计算放射增敏比;用流式细胞术检测4Gy照射后细胞凋亡率;用蛋白印迹实验检测凋亡相关蛋白cleaved PARP-1、cleaved caspase-3及DNA损伤相关蛋白γ-H2AX表达水平。[结果]转染siDYRK1B的ECA109细胞中DYRK1B mRNA和蛋白水平明显下降(P<0.05)。与对照组相比,转染siDYRK1B的ECA109细胞放射处理后细胞存活减少,细胞凋亡率增加(P<0.05),细胞中cleaved PARP-1、cleaved caspase-3、γ-H2AX蛋白表达水平升高(P<0.05)。siDYRK1B转染的ECA109细胞放射增敏比为1.711。[结论]下调DYRK1B明显增强食管鳞癌细胞的放射敏感性,其机制可能与促进细胞凋亡、抑制DNA损伤修复有关。

[Purpose] To investigate the effect of DYRK1B on radiosensitivity of esophageal squamous cell carcinoma and its potential mechanism. [Methods] Human esophageal squamous cell carcinoma ECA109 cells were transfected with siNC and siDYRK1B by Lipofectamine 3000,respectively. The expression levels of DYRK1B in ECA109 cells were detected by RT-PCR and Western blot 48 h after transfection. The transfected ECA109 cells were exposed to ionizing radiation,the survival of cells after radiation was evaluated by colony formation assay and the sensitive enhancement ratio(SER) was calculated accordingly. Apoptosis rate after 4Gy irradiation was detected by flow cytometry;the expression levels of cleaved PARP-1,cleaved caspase-3 and γ-H2AX were detected by Western blot. [Results] DYRK1B mRNA and protein levels were significantly decreased in ECA109 cells transfected with siDYRK1B(P<0.05). When treated with ionizing radiation,the survival of ECA109 cells transfected with siDYRK1B was decreased(P<0.05),the apoptosis rate was increased(P<0.05),and the expression levels of cleaved PARP-1,cleaved caspase-3,and γ-H2AX proteins were upregulated compared with the control group(P<0.05). The SER in ECA109-siDYRK1B cells was 1.711. [Conclusion] Down-regulation of DYRK1B significantly enhances the radiosensitivity of esophageal squamous cell carcinoma ECA109 cells,which may be related to promoting apoptosis and inhibiting DNA damage repair.