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目的:探讨电离辐射对人食管鳞癌TE-1细胞株的上皮间质转化(epithelial-mesenchymal transition,EMT)的影响和潜在机制。方法:用不同强度的电离辐射(0,2,4,8 Gy)处理TE-1细胞,定量PCR检测Snail 1、激活蛋白1(activator protein-1,AP-1)的mRNA表达,蛋白质印迹法检测Snail 1、Slug、E-钙黏蛋白、波形蛋白的表达量,免疫荧光观察E-钙黏蛋白、波形蛋白的表达,采用划痕实验和Transwell实验检测细胞迁移能力;结合Notch通路抑制剂DAPT,经蛋白质印迹法测定电离辐射(4 Gy)后TE-1细胞中Notch 1/NICD/Hes 1通路蛋白、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白表达变化。结果:电离辐射处理后TE-1细胞EMT相关转录因子Snail 1、Slug、AP-1明显上调,E-钙黏蛋白的表达显著减少,波形蛋白、MMP-9的表达显著增多,且4Gy处理水平即具有明显的促进作用(P<0.01);电离辐射上调了TE-1细胞的迁移能力(P<0.05);电离辐射通过上调Notch 1/NICD/Hes 1信号通路蛋白的表达水平,促使EMT相关蛋白的上调(P<0.01)。结论:电离辐射通过人食管鳞癌TE-1细胞中Notch信号通路诱导EMT及迁移。
Objective: To investigate the effect and potential mechanism of ionizing radiation on epithelial-mesenchymal transition (EMT) in human esophageal squamous cell carcinoma TE-1 cell line. Methods: TE-1 cells were treated with different intensities of ionizing radiation (0, 2, 4, 8 Gy). The mRNA expressions of Snail 1 and AP-1 were detected by quantitative PCR. Western blotting The expression of Snail 1, Slug, E-cadherin and vimentin were detected by immunohistochemistry. The expression of E-cadherin and vimentin was detected by immunofluorescence. The migration ability of cells was detected by scratch assay and Transwell assay. Combined with Notch pathway inhibitor DAPT The protein expression of Notch 1 / NICD / Hes 1 pathway and matrix metalloproteinase-9 (MMP-9) in TE-1 cells after ionizing radiation (4 Gy) was measured by Western blotting. Results: The EMT-related transcription factors Snail 1, Slug and AP-1 in TE-1 cells were significantly increased, the expression of E-cadherin was significantly decreased, the expression of vimentin and MMP-9 was significantly increased after treated with ionizing radiation (P <0.01). Ionizing radiation up-regulated the migration ability of TE-1 cells (P <0.05). Ionizing radiation induced EMT by up-regulating the expression of Notch 1 / NICD / Hes 1 signaling pathway proteins Protein upregulation (P <0.01). Conclusion: Ionizing radiation induces EMT and migration through the Notch signaling pathway in human esophageal squamous cell carcinoma TE-1 cells.