目的 探讨辐射表达载体Egr1-survivin shRNA联合放疗对食管鳞癌ECA109细胞凋亡及放射敏感性的影响。方法 将合成的辐射表达载体Egr1-survivin shRNA经LipofectamineTM2000转染ECA109细胞后给予放疗,并以YM155联合放疗为主要对照,对比通过qPCR、Western-Blot检测处理后24 h各组细胞survivin的mRNA及蛋白表达,用流式细胞仪检测处理后48 h细胞周期及凋亡率变化。结果 ECA109细胞经Egr1-survivin shRNA质粒转染联合放疗处理后,survivin mRNA及蛋白的表达量不但明显低于空白对照组和空载体对照组,也明显低于YM155处理联合放疗。经Egr1-survivin shRNA联合放疗处理后,ECA109细胞的G2与S期细胞比例明显增加,产生明显G2与S期阻滞;凋亡率明显增高,高于YM155联合放疗组,更明显高于空白对照组和空载体对照组,差异均有统计学意义(P＜0.01)。结论 辐射表达载体Egr1-survivin shRNA联合放疗可进一步增加食管鳞癌ECA109细胞凋亡,增强其放射敏感性,是一种值得探索的辅助放疗途径。
Abstract
Objective To investigate the effect of radiative expression vector Egr1-survivin shRNA combined with irradiation on the apoptosis and radiotherapy sensitivity of esophageal squamous cell carcinoma ECA109 cells. Methods The radiative expression vector Egr1-survivin shRNA was designed and transfected into ECA109 cells by LipofectamineTM2000 before radiotherapy was started. The treatment with YM155 combined with irradiation was established as the main control. The expression levels of survivin mRNA and protein in each treatment group were detected 24 hours after treatment with qPCR and Western Blot respectively. The cell cycles and cell apoptosis of each treatment group were detected by flow cytometry 48 hours after treatment respectively. Results In ECA109 cells, the expression levels of survivin mRNA and protein were more significantly down-regulated in transfected Egr1-survivin shRNA plasmid combined with irradiation group than in YM155 combined with irradiation group, the blank control group, and the blank vector control group. Compared with the last three groups, G2 and S cycle cell rates were significantly increased, while the apoptosis rate was clearly decreased in Egr1-survivin shRNA combined with irradiation group (P<0.01) respectively). Conclusions The radiative expression vector Egr1-survivin shRNA combined with irradiation can promote the apoptosis of ECA109 cells and enhance their radiotherapy sensitivity. It promises to be a therapeutic method in conjunction with radiotherapy.
关键词
食管鳞癌 /
Egr1 /
survivin /
放射敏感性 /
基因-放射治疗
Key words
esophageal squamous cell carcinoma /
Egr1 /
survivin /
radiosensitivity /
gene-radiotherapy
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基金
新疆维吾尔自治区自然科学基金(2016D01C345)
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