敲低额外11-19白血病融合基因蛋白通过激活线粒体通路诱导非小细胞肺癌A549细胞凋亡  

Knockdown of extra eleven-nineteen leukemia fusion gene protein induces apoptosis of non-small cell lung cancer A549 cells by activating mitochondrial pathway

作　　者：周铮铮 徐凯 陈义钢[2] 孙庆科 Zhou Zhengzheng;Xu Kai;Chen Yigang;Sun Qingke(Department of Oncology,Wuxi No.8 People’s Hospital,Wuxi 214000,China;Department of General Surgery,Wuxi Second People’s Hospital,Wuxi 214000,China;Thoraco-Cardiac Surgery,Chinese People’s Liberation Army Rocket Force Characteristic Medical Center,Beijing 100088,China)

机构地区：[1]无锡市第八人民医院肿瘤科,无锡214000 [2]无锡市第二人民医院普外科,无锡214000 [3]火箭军特色医学中心胸心外科,北京100088

出　　处：《中国组织化学与细胞化学杂志》2023年第4期385-391,共7页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的 探讨额外11-19白血病融合基因蛋白(extra eleven-nineteen leukemia fusion gene protein, EEN)对非小细胞肺癌(non-small cell lung cancer, NSCLC)的影响及其作用机制。方法 用免疫组织化学染色检测EEN在NSCLC组织和癌旁组织的表达水平,Western blot检测EEN在正常肺上皮细胞BEAS-2B细胞和NSCLC细胞A549细胞中的水平。在A549细胞中转染sh-EEN后,使用MTT测定细胞活性,Annexin V-FITC/PI流式细胞术和TUNEL染色分析细胞凋亡,使用荧光探针DCFH-DA测量细胞内ROS生成水平,使用JC-1染色测定线粒体膜电位的变化,通过Western blot分析凋亡相关蛋白水平。结果 与癌旁组织相比,EEN在NSCLC组织中表达上调;与BEAS-2B细胞相比,EEN在A549细胞中水平明显升高。沉默A549细胞EEN后,A549细胞生长受到抑制,细胞凋亡显著增加,ROS水平升高,线粒体膜电位丢失,Cyt C释放入细胞质,同时伴随着Bcl-2降低、Bax升高和Caspase-3活化。结论 EEN在NSCLC中表达增加。敲低EEN可诱导NSCLC细胞凋亡,且这一过程至少涉及到ROS/线粒体凋亡信号通路的激活。Objective To investigate the impact and mechanism of action of the extra eleven-nineteen leukemia fusion gene protein(EEN)on non-small cell Lung cancer(NSCLC).Methods Immunohistochemical staining was used to detect the expression levels of EEN in NSCLC tissues and adjacent non-cancerous tissues,while Western blot was employed to measure the levels of EEN in normal lung epithelial cells(BEAS-2B)and NSCLC cells(A549).After transfecting A549 cells with sh-EEN,cell viability was assessed using the MTT assay,apoptosis was analyzed through Annexin V-FITC/PI flow cytometry and TUNEL staining.The generation of intracellular ROS was measured using the fluorescent probe DCFH-DA,changes in mitochondrial membrane potential were determined by JC-1 staining,and the levels of apoptosis-related proteins were examined via Western blot analysis.Results EEN expression was upregulated in NSCLC tissues compared to adjacent non-cancerous tissues,and levels of EEN were significantly higher in A549 cells than in BEAS-2B cells.Silencing EEN in A549 cells inhibited cell growth,significantly increased apoptosis,raised ROS levels,led to loss of mitochondrial membrane potential,and resulted in the release of Cyt C into the cytoplasm.This was accompanied by a decrease in Bcl-2,an increase in Bax,and activation of Caspase-3.Conclusion EEN expression is increased in NSCLC.Knocking down EEN can induce apoptosis in NSCLC cells,and this process involves at least the activation of the ROS/mitochondrial apoptosis signaling pathway.

关 键 词：非小细胞肺癌 额外11-19白血病融合基因蛋白 细胞凋亡 

分 类 号：R365[医药卫生—病理学]