【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。更多还原

【Abstract】 Objective To investigate whether Notch1 gene to changes the proliferation of human neuroblastoma SH-SY5Y cell line.Methods Notch1 gene activation of SH-SY5Y cells was blocked using γ-secretase inhibiter DAPT or knockdown with recombination lentivirus which express Notch1-shRNA.RT-PCR and Western Blot were applied to monitor the validity of down-regulation of notch1 and Hes1 expression.MTT assay was performed for examining cell proliferation in vitro.The influence of Notch1 RNAi on the implanted tumor growth was observed.ResultsDAPT and the lentiviral vectors expressing Notch1-shRNA were efficient in down-regulating notch1(NICD) and Hes1 expression.The down-regulation of Notch1 gene inhibited cell proliferation significantly(P<0.01).The implanted tumor weight in Notch1-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.更多还原