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作 者:刘学良[1] 曾泉[1] 邱志利[1] 胡国华[1]
机构地区:[1]重庆医科大学附属第一医院耳鼻咽喉科,重庆400016
出 处:《重庆医科大学学报》2013年第8期864-867,共4页Journal of Chongqing Medical University
基 金:重庆市自然科学基金重点资助项目(编号:cstc2012jjB10015);重庆市卫生局资助项目(编号:cstc2010220)
摘 要:目的:研究抑制NFBD1表达后,对鼻咽癌(nasopharyngeal carcinoma,NPC)细胞株HNE-1凋亡的影响。方法:用NFBD1shRNA慢病毒感染HNE-1,采用real-time PCR和Western blot检测NFBD1 shRNA慢病毒对NFBD1的抑制效果;Hoechst33342染色观察细胞核的改变并结合流式细胞仪(flow cytometry,FCM),TUNEL法检测细胞的凋亡情况。结果:NFBD1 shRNA慢病毒在mRNA和蛋白质水平均能高效、特异性地抑制NFBD1的表达;Hoechst33342染色结果显示,在NFBD1下调组,呈现固缩且边缘化的细胞核数量明显增加(χ2=5.367;P=0.021);TUNEL染色结果显示,在NFBD1下调组,显现荧光的凋亡细胞明显增加,FCM检测结果显示,在NFBD1下调组,细胞凋亡率明显增加(χ2=326.86;P<0.001)。结论:抑制NFBD1基因的表达后,能有效促进HNE-1凋亡,这为寻求NPC的治疗靶点提供新的思路。Objective:To investigate the effects of nuclear factor with B RCT domains protein 1 (NFBD1)gene silencing on apoptosis in nasopharyngeal carcinoma(NPC) cells. Methods:Retroviruses expressing NFBD1 shRNA were applied to infect NPC cell line HNE-1. Real-time PCR and Western bolt were used to confirm the inhibitory effects of retroviruses against NFBD1. Flow cytometry(FCM), TUNEL and Hoechest33342 staining methods were used to tested cell apoptosis. Results :NFBD1 shRNA retrovirus can inhibit mRNA and protein expressions of NFBD1 efficiently and specifically. According to the results of Hoechst33342 staining, nucleus exhibiting pycnosis and marginalization were increased significantly in experimental group than in control group(x2=5.367,P=0.021). Accord- ing to the results of TUNEL staining, apoptotic cells presenting fluorescence were increased significantly in experimental group than in control group. According to the results of FCM, cell apoptosis was significantly increased in experimental group than in control group (X2=326.g6,P〈0.001). Conclusions:NFBD1 gene silencing in nasopharyngeal carcinoma cell line HNE-1 can effectively promote the HNE-1 apoptosis,which provide new ideas for treatment of NPC.
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