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

作　　者：孙为增[1] 林国雄[1] 林海[1] 

机构地区：[1]海南省人民医院,海口570311

出　　处：《山东医药》2017年第40期22-25,共4页Shandong Medical Journal

基　　金：海南省自然科学基金资助项目(20168283)

摘　　要：目的探讨沉默钙连蛋白(Calnexin)对肾母细胞瘤细胞凋亡及内质网应激、JNK信号通路的影响。方法将肾母细胞瘤SK-NEP-1细胞分为对照组、空载体组、实验组。对照组正常培养不转染。空载体组转染空白质粒,实验组转染Calnexin shRNA。转染24 h后全部更换为正常培养液继续培养48 h。采用实时荧光定量PCR法检测各组Calnexin mRNA表达,TUNEL法检测细胞凋亡指数(AI),Western blotting法检测葡萄糖调节蛋白78(GRP78)、需肌醇酶1(IRE1)、肿瘤坏死因子受体相关因子2(TRAF2)、凋亡信号调节激酶1(ASK1)、磷酸化凋亡信号调节激酶1(p-ASK1)、c-Jun N末端激酶(JNK)及磷酸化c-Jun N末端激酶(p-JNK)蛋白相对表达量。结果 Calnexin mRNA相对表达量:实验组、空载体组、对照组分别为0.28±0.01、1.01±0.06、1.00±0.08;实验组低于空载体组和对照组(P<0.05),但空载体组与对照组比较无统计学差异(P>0.05)。AI:实验组、空载体组、对照组分别为32.86±3.72、5.46±0.68、5.26±0.17;实验组高于空载体组和对照组(P<0.05),但空载体组与对照组比较无统计学差异(P>0.05)。GRP78、IRE1、TRAF2、ASK1、p-ASK1、JNK及p-JNK蛋白相对表达量:实验组高于空载体组和对照组(P均<0.05),但空载体组与对照组比较无统计学差异(P均>0.05)。结论沉默Calnexin可引发内质网应激并激活JNK细胞凋亡通路,进而促进肾母细胞瘤SK-NEP-1细胞凋亡。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＆lt;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 be

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 

分 类 号：R737.11[医药卫生—肿瘤]