摘要
目的探讨转染人Notchl受体胞内段(NIcD)质粒激活Notchl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-timePcR检测靶基因Hesl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;caspase3试剂盒检测NICD转染后凋亡通路关键酶caspase3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Hesl表达,提示激活Notchl信号通路。NICD转染可显著促进胰腺癌细胞生长增殖;caspase3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Notchl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。
Objective To observe the effect of activation of Notch1 signaling pathway by Notch intraceUular 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 Hesl 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. Hesl 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.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第10期1494-1498,共5页
Journal of Southern Medical University
基金
中央高校基本科研业务(2011SCU11050)
四川省科技支撑计划(2011SZ0293
2011SZ0087)
关键词
胰腺癌
NOTCH信号通路
Notch胞内段
增殖
凋亡
pancreatic cancer
Notch signaling pathway
Notch intracellular domain
proliferation
apoptosis