摘要
目的探讨RNA干扰核因子E2相关因子2(Nrf2)对食管癌细胞生物学行为的影响。方法建立RNA干扰下调Nrf2表达的Eca-109食管癌细胞(Si-Nrf2),并构建阴性对照组细胞(Si-control)。MTT法、平板克隆形成实验、流式细胞术、Transwell小室实验及Western blotting法检测下调Nrf2表达对食管癌细胞增殖、凋亡及侵袭转移、细胞放射敏感性及相关蛋白表达的影响。结果转染48h后,Si-Nrf2细胞中Nrf2蛋白相对表达量为0.209±0.013,Si-control细胞中则为0.852±0.077,两者差异具有统计学意义(P<0.05)。与Si-control细胞比较,下调Nrf2表达水平后的Si-Nrf2细胞的增殖能力明显减弱、细胞凋亡增多、侵袭转移能力降低,放疗敏感性增高,血红素加氧酶(HO-1)、抗凋亡蛋白Bcl-2、基质金属蛋白酶-9(MMP-9)蛋白的表达量明显下调(P<0.05)。结论 Nrf2可能通过调控HO-1、Bcl-2及MMP-9蛋白表达参与食管癌细胞增殖、凋亡、侵袭转移及放疗敏感性这一生物学过程。
Objective To investigate the effect of down-regulation of nuclear factor erythroid 2-related factor（ Nrf2） by RNAi on biological behaviors in esophageal cancer cells. Methods Eca-109 cell line which was down-regulated Nrf2 by RNAi（ Si-Nrf2） and negative control cell（ Si-control） were established. Si-Nrf2 cells and Si-control cells were transfected by Lipofectamine 2000. Cell proliferation,apoptosis,cell metastasis and relative protein expression were analyzed by MTT assay,colony formation assay,flow cytometry,transwell and western blotting method after transfection. Results Western blotting showed that the expression of Nrf2 in the SiNrf2 cells was 0. 209 ± 0. 013,which was significantly lower than that in the Si-control cells（ 0. 852 ± 0. 077,P 〈 0. 05）. The result of biological function shown that Eca-109 cell down-regulated Nrf2 had a lower survival fraction,higher cell apoptosis,and significant decrease in migration and invasion,higher radiosensitivity and lower expression of HO-1,Bcl-2,MMP-9 protein compared with those of Si-control cells（ P 〈 0. 05）. Conclusion Nrf2 may involve biological process of cell proliferation,apoptosis,invasion and metastasis and radiation sensitivity in esophageal cancer by regulating the expression of HO-1,Bcl-2 and MMP-9 protein.
出处
《临床肿瘤学杂志》
CAS
2014年第12期1057-1061,共5页
Chinese Clinical Oncology
基金
河北省科技支撑计划资助项目(112761162)
关键词
NRF2
食管鳞癌
细胞增殖
细胞凋亡
Nrf2
Esophageal squamous cell carcinoma
Cell proliferation
Cell apoptosis
