CKS1表达与食管癌细胞辐射敏感性的关系研究

Effects of CKS1 Expression on Radio-sensitivity of Esophageal Carcinoma

目的:探讨CKS1表达对食管癌细胞辐射敏感性的影响,初步研究其分子机理。方法:用Western-blotting方法筛选CKS1低表达和高表达的食管癌细胞系;构建CKS1正义表达载体p-pcDNA 3.1/myc-His A-CKS1和RNA干扰载体CKS1 siRNA,分别转染CKS1低表达细胞和高表达细胞,用不同剂量γ-射线照射各组细胞,克隆形成实验检测细胞辐射敏感性的差异。结果:CKS1在四种食管癌细胞中的表达水平依次为EC9706>KYSE510>KYSE450>KYSE150。用p-pcDNA 3.1/myc-His A-CKS1表达载体转染KYSE150细胞后CKS12表达升高,不同剂量γ-射线照射后细胞的克隆形成能力显著高于母系对照组(P<0.01)。RNA干扰载体转染KYSE510细胞后CKS1表达水平降低,不同剂量γ-射线照射后细胞的克隆形成能力显著低于母系对照组(P<0.01)。敲降CKS1表达后DNA损伤修复相关蛋白RAD51表达下降,KU70表达没有变化。CKS1过表达后RAD51表达升高,KU70表达没有变化。结论:CKS1表达与食管癌细胞的辐射敏感性密切相关,可能通过影响DNA损伤修复发挥作用。

Objective：To explore the correlation of CKS1 expression with radio-sensitivity of Esophageal Carcinoma and its mechanism.Methods：Detecting the level of CKS1 in four different Esophageal carcinoma cell lines using western-blotting.Constructing p-pcDNA 3.1/myc-His A-CKS1 expression and CKS1 siRNA vector and transfecting CKS1 low or high expression cell,respectively.Clone forming assay was used to detect the cell proliferation ability of different groups after different doses of γ-ray irradiation.Results：The sequence of CKS1 expression level in four different Esophageal Carcinoma cell line was EC9706KYSE510KYSE450KYSE150.Compared with parent control cells,the clone forming ability of CKS1 high expression cells was significantly increased（P0.01）.In contrast,the clone forming ability of KYSE510 cell was significantly decreased after CKS1 knock-down（P0.01）.RAD51 expression was decreased or increased after CKS1 knock-down or overexpression,respectively.But KU70 expression did not change.Conclusion：CKS1 may have effect on radio-sensitivity of Esophageal Carcinoma via DNA damage pathway.