沉默NFBD1表达对鼻咽癌细胞凋亡的影响

Effects of NFBD1 gene silencing on apoptosis in nasopharyngeal carcinoma cells

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NFBD1 shRNA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。

Abstract:

Objective：To investigate the effects of nuclear factor with BRCT 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（ χ2=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（ χ2=326.86，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.