下调Notch1信号抑制人神经母细胞瘤SH-SY5Y的增殖能力  

Down-regulation of Notch-1 signaling contributes to cell growth inhibition in human neuroblastoma SH-SY5Y cells

作　　者：郑志竑[1] 林玲[1] 张燕[1,2] 郑莺凤[1,3] 

机构地区：[1]福建医科大学基础医学院生物化学与分子生物学系,福建福州350004 [2]杭州艾迪康医学检验中心 [3]加拿大Manitoba大学

出　　处：《基础医学与临床》2013年第5期578-583,共6页Basic and Clinical Medicine

基　　金：福建省自然科学基金(C0810017);福建医科大学研究基金(JS06008)

摘　　要：目的探讨Notch1基因与人神经母细胞瘤SH-SY5Y细胞增殖能力的关系。方法应用-分泌酶抑制剂DAPT或表达Notch1-shRNA慢病毒载体阻断SH-SY5Y细胞Notch1信号的活化,RT-PCR和Western blot法行Notch1及Hes1表达水平的检测,MTT法和裸鼠体内种植检测瘤细胞体内外增殖能力的变化。结果 DAPT处理和表达Notch1-shR-NA慢病毒载体转染均能减少细胞内NICD和Hes1表达,其细胞体外增殖能力受到明显抑制(P<0.01),Notch1 RNAi细胞体内种植瘤重为0.20±0.13 g,明显轻于对照组的0.89±0.58 g(P<0.01)。结论 Notch1信号的活化与人神经母细胞瘤SH-SY5Y细胞的增殖能力密切相关,有望成为神经母细胞瘤基因治疗的潜在靶点。Objective To SYSY cell line. Methods investigate whether Notchl gene to changes the proliferation of human neuroblastoma SH- Notchl gene activation of SH-SY5Y cells was blocked using γ-secretase inhibiter DAPT or knockdown with recombination lentivirus which express Notchl-shRNA. RT-PCR and Western Blot were applied to monitor the validity of down-regulation of notchl and Hesl expression. MTT assay was performed for examining cell proliferation in vitro. The influence of Notch1 RNAi DAPT and the lentiviral vectors expressing Notchl-shRNA on the implanted tumor growth was observed. Results were efficient in down-regulating notchl （NICD） and Hesl expression. The down-regulation of Notch1 gene inhibited cell proliferation significantly （P 〈 0. 01 ）. The im- planted tumor weight in Notchl-knockdown group was markedly lower than that in control group （0. 20 ± 0. 13 g vs 0. 89± 0.58 g, P 〈 0. 01 ）. Conclusions Notch1 gene activation, which can lead to the change of proliferation of SH-SY5Y cell line, is a potential target molecule for neuroblastoma of gene therapy.

关 键 词：神经母细胞瘤 NOTCH1基因 RNA干扰 DAPT 细胞增殖 

分 类 号：R739.4[医药卫生—肿瘤]