南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (10): 1494-.

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激活Notch1信号通路对胰腺癌细胞增殖的影响

杜潇,张思琴,程中,李旸,王自强,陈志新,胡建昆,周总光   

  • 出版日期:2013-10-20 发布日期:2013-10-20

Effect of Notch1 signaling pathway activation on pancreatic cancer cell proliferation in vitro

  • Online:2013-10-20 Published:2013-10-20

摘要: 目的探讨转染人Notch1受体胞内段(NICD)质粒激活Notch1信号通路对胰腺癌细胞增殖的影响及其可能机制。方法构
建带免疫荧光的NICD质粒与对照质粒,细胞转染后荧光激发显微镜下观察转染效率,Real-time PCR检测靶基因Hes1表达水
平变化;CCK-8法计算NICD质粒转染后胰腺癌细胞增殖的影响;caspase 3试剂盒检测NICD转染后凋亡通路关键酶caspase 3
活性的变化。结果荧光激发显示NICD转染效率在60%~80%之间;NICD转染可促进靶基因Hes1表达,提示激活Notch1信号
通路。NICD转染可显著促进胰腺癌细胞生长增殖;caspase 3活性在NICD转染后明显下降,差异均具有统计学意义。结论应
用NICD质粒转染激活Notch1信号通路活性可通过抑制凋亡而促进胰腺癌细胞生长增殖。

Abstract: Objective To observe the effect of activation of Notch1 signaling pathway by Notch intracellular domain (NICD)
plasmid transfection on pancreatic cancer cell proliferation and explore the underlying mechanism. Methods The transfection
rates were observed under microscope with fluorescence stimulation, and mRNA expression levels of Hes1 were detected by
real-time PCR. Cell proliferation changes were evaluated by CCK-8 after NICD and control plasmid transfection in pancreatic
cancer cells. Caspase 3 activity was examined using a caspase 3 detection kit. Results The transfection rates of NICD plasmid
were up to 80% by fluorescence stimulation observation. Hes1 expression was significantly increased after NICD plasmid
transfection, suggesting the activation of Notch1 signaling pathway. NICD plasmid transfection significantly promoted cancer
cell proliferation compared to control plasmid transfeciton. The activities of caspase 3 were obviously decreased after NICD
plasmid transfection in 3 pancreatic cancer cell lines. Conclusions Activation of Notch1 signaling pathway by NICD plasmid
transfection can promote the proliferation of pancreatic cancer cells by inhibiting the apoptosis pathway.