靶向沉默Notch活化复合酶NACK对神经母细胞瘤SK-N-BE(2)细胞增殖、凋亡的影响  

Effects of targeted silencing Notch activated complex kinase NACK on proliferation and apoptosis of neuroblastoma SK-N-BE(2)cells

作　　者：刘建英[1] 于欣欣[2] 王建勇 李爱敏[1] 李林林 郝晓昱 LIU Jianying;YU Xinxin;WANG Jianyong;LI Aimin;LI Linlin;HAO Xiaoyu(Department of Pediatrics, Yantai Yuhuangding Hospital, Medical College of Qingdao University, Yantai 264000, China;Department of Pediatrics, People′s Hospital of Binzhou City, Binzhou 256610,China)

机构地区：[1]青岛大学医学院附属烟台毓璜顶医院儿科,烟台264000 [2]滨州市人民医院儿科,滨州256610

出　　处：《中国小儿血液与肿瘤杂志》2022年第2期90-96,共7页Journal of China Pediatric Blood and Cancer

基　　金：烟台市科技计划项目(201625)。

摘　　要：目的探讨靶向沉默Notch活化复合酶NACK对神经母细胞瘤SK-N-BE(2)细胞增殖及凋亡的影响,并探讨其可能的作用机制。方法通过脂质体介导siRNA转染SK-N-BE(2)细胞,qRT-PCR、Western blot方法检测转染后NACK基因的沉默效率;CCK-8法检测各组细胞增殖抑制率;流式细胞术检测各组细胞凋亡率;qRT-PCR、Western blot方法检测靶向抑制NACK表达后Notch1信号传导通路下游靶基因Hes1、c-Myc和n-Myc的表达情况。结果成功转染NACK siRNA的实验组细胞的NACK mRNA基因及其蛋白表达水平明显降低(P<0.05)。CCK-8法检测到实验组细胞增殖率较阴性和空白对照组相比明显下降(P<0.01),流式细胞术检测到实验组细胞凋亡率较阴性和空白对照组明显增加(P<0.01)。Western blot、qRT-PCR方法检测到实验组Notch1信号通路下游相关靶基因Hes1、c-Myc和n-Myc的蛋白水平和mRNA表达水平较阴性对照组及空白对照组均明显下降,差异有统计学意义(P<0.05)。结论靶向沉默NACK基因可以抑制神经母细胞瘤SK-N-BE(2)细胞增殖,促进凋亡,有望成为神经母细胞瘤基因治疗的潜在靶点。Objective To investigate the effects of targeted silencing Notch activated complex kinase(NACK)on proliferation and apoptosis of SK-N-BE(2)cells and to explore its possible mechanism.Methods The SK-N-BE(2)cells were transfected by liposome-mediated siRNA,qRT-PCR and Western blot methods were used to detect the silencing efficiency of the NACK gene after transfection,CCK-8 method was used to detect the cell proliferation inhibition rate of each group and flow cytometry was used to analyze the apoptosis rate of each group.The qRT-PCR and Western blot methods were used to detected the expressions of the downstream target genes Hes1,c-Myc and n-Myc of Notch1 signaling pathway after NACK silenced.Results The expression levels of NACK gene mRNA and protein of cells which successfully transfected by NACK siRNA were significantly reduced(P<0.05).CCK-8 method showed that the cell proliferation in the experimental group was significantly decreased compared with the negative control group and the blank control group.The flow cytometry showed that the apoptosis in the experimental group was significantly increased compared with the negative control group and the blank control group(P<0.01).Western blot and qRT-PCR methods showed that the protein and mRNA expression levels of the downstream target genes Hes1,c-Myc,and n-Myc of the Notch1 signaling pathway of the experimental group were significantly lower than the negative control group and the control group.The difference was statistically significant(P<0.05).Conclusions Targeted silencing NACK gene can inhibit the proliferation and promote apoptosis of neuroblastoma SK-N-BE(2)cells,and NACK is expected to be a potential target of neuroblastoma gene therapy.

关 键 词：神经母细胞瘤 NOTCH NACK 细胞增殖 凋亡 

分 类 号：R73[医药卫生—肿瘤]