山东医药2017，Vol.57Issue(40)：22-25,4.DOI:10.3969/j.issn.1002-266X.2017.40.006
沉默钙连蛋白对人肾母细胞瘤细胞凋亡及内质网应激、JNK信号通路的影响
Effects of silencing calnexin on apoptosis, endoplasmic reticulum stress, and JNK signaling pathway of human nephroblastoma cells
摘要
Abstract
Objective To investigate the effects of silencing calnexin on the apoptosis , endoplasmic reticulum stress , and JNK signaling pathway of human nephroblastoma cells .Methods The nephroblastoma SK-NEP-1 cells were divided into the control group , empty vector group , and experimental group .The cells in the control group were not transfected . The cells in the empty vector group were transfected with the blank plasmid , and the experimental group with Calnexin shR-NA.After 24-hour transfection, all were replaced with normal culture medium and then were cultured for 48 h.The expres-sion of Calnexin mRNA was detected by real-time fluorescence quantitative PCR .The apoptotic index ( AI) was detected by TUNEL method.Western blotting was used to detect the relative expression of glucose-regulated protein 78 (GRP78), ino-sitol-requiring enzyme 1 (IRE1), tumor necrosis factor receptor-associated factor 2 (TRAF2), apoptosis signal-regulating kinase 1 (ASK1), p-ASK1, c-Jun N-terminal kinase (JNK) and phosphorylated c-Jun N-terminal kinase (p-JNK) pro-tein.Results The relative expression of calnexin mRNA was 0.28 ±0.01, 1.01 ±0.06, and 1.00 ±0.08 in the experi-mental group , empty vector group , and control group , respectively .The experimental group was lower than the empty vec-tor group and the control group (P<0.05), but there was no significant difference between the empty vector group and the control group (P>0.05).AI in the experimental group, empty vector group, and control group were 32.86 ±3.72, 5.46 ±0.68, and 5.26 ±0.17; the experimental group was higher than the empty vector group and the control group ( P<0.05), but there was no significant difference between the empty vector group and the control group (P>0.05).The rela-tive expression levels of GRP78, IRE1, TRAF2, ASK1, p-ASK1, JNK and p-JNK were higher in the experimental group than those in the empty vector group and the control group (all P<0.05), but there was no significant difference between the empty vector group and the control group (P>0.05).Conclusion Silencing calnexin can lead to endoplasmic reticu-lum stress and activate JNK cell apoptosis pathway , and thus promote the apoptosis of nephroblastoma SK-NEP-1 cells.关键词
肾母细胞瘤/钙连蛋白/细胞凋亡/内质网应激/JNK信号通路Key words
nephroblastoma/calnexin/apoptosis/endoplasmic reticulum stress/JNK signaling pathway分类
医药卫生引用本文复制引用
孙为增,林国雄,林海..沉默钙连蛋白对人肾母细胞瘤细胞凋亡及内质网应激、JNK信号通路的影响[J].山东医药,2017,57(40):22-25,4.基金项目
海南省自然科学基金资助项目(20168283). （20168283）
