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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 investigate the effect of CKS1 expression on the radiosensitivity of esophageal cancer cells and its molecular mechanism. Methods: The esophageal cancer cell lines with low expression and high expression of CKS1 were screened by Western-blotting. CKS1 positive expression vector p-pcDNA3.1 / myc-His A-CKS1 and RNA interference vector CKS1 siRNA were constructed and transfected into CKS1 low expression cells And high expression of cells, with different doses of γ-ray irradiation of each group of cells, colony formation assay to detect differences in cell radiation sensitivity. Results: The expression level of CKS1 in four esophageal cancer cells was EC9706> KYSE510> KYSE450> KYSE150. The expression of CKS12 in KYSE150 cells transfected with p-pcDNA3.1 / myc-His A-CKS1 expression vector was significantly higher than that in the maternal control group (P <0.01) after irradiation with different doses of γ-rays. The expression level of CKS1 in KYSE510 cells transfected with RNA interference vector was decreased. The colony formation ability of cells after γ-ray irradiation with different doses was significantly lower than that of maternal control group (P <0.01). After knocking down CKS1 expression, the expression of DNA damage repair related protein RAD51 decreased, KU70 expression did not change. After overexpression of CKS1, the expression of RAD51 was up-regulated and the expression of KU70 did not change. Conclusion: The expression of CKS1 is closely related to the radiosensitivity of esophageal cancer cells and may play a role in the DNA damage repair.