Autophagy inhibition contributes to radiation sensitization of esophageal squamous carcinoma cells
Autophagy inhibition contributes to radiation sensitization of esophageal squamous carcinoma cells
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DOI：
10.1111/j.1442-2050.2010.01156.x
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发表时间：
2011-08-01
影响因子：
2.6
通讯作者：
Fu, T.
中科院分区：
文献类型：
--
作者：
Chen, Y. S.;Song, H. X.;Fu, T.
放射治疗是食道癌治疗策略中有用的组成部分。研究了自噬在人食道鳞癌细胞对电离辐射的反应中的作用。采用细胞存活率和克隆形成法检测自噬抑制物(3-MA)对食管鳞癌细胞的放射敏感性。流式细胞仪检测细胞凋亡率和细胞周期分析；DAPI染色检测细胞凋亡率。免疫印迹法检测BECLIN-1和LC3的表达。在电子显微镜下进行超微结构分析。6Gy射线照射可诱导TE-1细胞自噬小体大量积聚，并明显上调Beclin-1和Lc3-II的表达。与单纯辐射相比，3-MA联合辐射可显著降低细胞存活率、自噬比率、BECLIN-1和LC3-II蛋白水平。抑制自噬可增加辐射诱导的细胞凋亡率和G2/M期细胞的比例。3-MA阻断自噬增强放射治疗对人食管鳞癌细胞的细胞毒作用提示抑制自噬可作为食管鳞癌的辅助治疗手段。
Radiotherapy is a useful component of treatment strategies for esophageal cancer. The role of autophagy in response to ionizing radiation was investigated in human esophageal squamous carcinoma cells. Cell viability and clonogenic survival assay were used to evaluate the radiosensitivity of autophagy inhibitor (3-MA) on esophageal squamous carcinoma cells. The percentage of apoptotic cells and cell cycle analysis were assessed by flow cytometry; DAPI staining was used to detect apoptotic cells. The expression of beclin-1 and LC3 was measured using a Western blot. The ultrastructural analysis was under the electron microscope. 6 Gy irradiation induced a massive accumulation of autophagosomes accompanied by strong upregulation of beclin-1 and LC3-II expression in TE-1 cells. Compared with radiation alone, 3-MA combined with radiation significantly decreased cell viability, as well as autophagic ratio, beclin-1, and LC3-II protein level. Inhibition of autophagy increased radiation-induced apoptosis and the percentage of G2/M-phase cells. Blockade of autophagy with 3-MA enhanced cytotoxicity of radiotherapy in human esophageal squamous carcinoma cells. It suggests that inhibition of autophagy could be used as adjuvant therapy to treat esophageal squamous cell carcinoma.