摘要
为探讨核孔蛋白88(nucleoporin 88,Nup88)在胃腺癌组织中的表达及对胃腺癌SGC-7901细胞增殖、凋亡的影响研究,用Western blotting检测收集胃腺癌组织中的Nup88。通过细胞转染siRNA Nup88和siRNA control抑制胃腺癌细胞中Nup88的表达。MTT检测转染后细胞增殖情况。流式细胞术检测转染后细胞凋亡情况。Western blotting检测细胞中NOTCH信号通路相关蛋白NICD1、NICD2、HES1和Cleaved Caspase-3水平。胃腺癌细胞与NOTCH信号通路抑制剂S2188作用后,检测细胞增殖和凋亡情况。结果显示Nup88在胃腺癌组织中的表达水平明显高于癌旁组织(P<0.01)。siRNA Nup88组细胞中Nup88水平明显低于siRNA control组(P<0.01)。siRNA Nup88组细胞存活率明显低于siRNA control组(P<0.01)。siRNA Nup88组细胞凋亡率明显高于siRNA control组(P<0.01)。siRNA Nup88组细胞中NICD1、NICD2、HES1水平明显低于siRNA control组(P<0.01)。siRNA Nup88组细胞中Cleaved Caspase-3水平明显高于siRNA control组(P<0.01)。抑制剂作用后的细胞增殖凋亡趋势与siRNA Nup88组一致。由此,Nup88在胃腺癌组织中表达上调。沉默Nup88后胃腺癌细胞增殖受到抑制,凋亡增多,作用机制与NOTCH信号通路有关。
To investigate the expression of Nup88 in gastric adenocarcinoma and its effect on the proliferation and apoptosis of gastric cancer cell line SGC-7901,the expression level of Nup88 was detected in gastric carcinoma by Western blotting.The inhibition of Nup88 expression in gastric adenocarcinoma cell line was carried out by transfection of siRNA Nup88 and siRNA control.The proliferation of transfected cells was detected by MTT.The apoptosis of transfected cells was detected by flow cytometry.Levels of NOTCH signal pathway related proteins including NICD1,NICD2,HES1 and Cleaved Caspase-3 in cells were detected by Western blotting.Proliferation and apoptosis of gastric cancer cells were examined after treatment by NOTCH signaling pathway inhibitor S2188.The results showed that Nup88 expression in gastric cancer tissues was significantly higher than that in adjacent tissues(P 0.01).Nup88 level in the siRNA Nup88 group was significantly lower than that in the siRNA control group(P 0.01).The cell survival rate of siRNA Nup88 group was significantly lower than that of the control siRNA group(P 0.01),while the apoptosis rate of siRNA Nup88 group was significantly higher than that of siRNA control group(P 0.01).The levels of NICD1,NICD2 and HES1 in siRNA Nup88 group were significantly lower than those in siRNA control group(P 0.01).The level of Cleaved Caspase-3 in siRNA Nup88 group was significantly higher than that in siRNA control group(P 0.01).The cell proliferation and apoptosis following the treatment by the inhibitor were similar to that of the siRNA Nup88 group.In conclusion,the expression of Nup88 is up-regulated in gastric adenocarcinoma.The proliferation of gastric cancer cells is inhibited,while apoptosis is increased after Nup88 knockdown,and the mechanism is related to NOTCH signaling pathway.
出处
《现代免疫学》
CSCD
北大核心
2017年第6期449-453,460,共6页
Current Immunology
基金
河北省卫计委青年科技课题(20150642)