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

发布时间：2019-06-27 13:10

【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(P0.05),但空载体组与对照组比较无统计学差异(P0.05)。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(P0.05),但空载体组与对照组比较无统计学差异(P0.05)。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。
[Abstract]:Objective To study the effect of calnexin on apoptosis and endoplasmic reticulum stress and JNK signaling pathway in renal cell tumor cells. Methods SK-NEP-1 cells were divided into control group, empty vector group and experimental group. The normal culture of the control group was not transfected. The empty vector group was transfected with a blank plasmid, and the experimental group was transfected with Calnexin shRNA. The expression of Calnexin mRNA in each group was detected by real-time fluorescence quantitative PCR, and the apoptosis index (AI) was detected by TUNEL method. The expression of glucose regulation protein 78 (GRP78), the desired inositol 1 (IRE1) and the tumor necrosis factor receptor-related factor 2 (TRAF2) were detected by the Western blotting method. The apoptosis signal regulates the relative expression of the kinase 1 (ASK1), the phosphorylated apoptosis signal modulating kinase 1 (p-ASK1), the c-Jun N terminal kinase (JNK), and the phosphorylated c-Jun N terminal kinase (p-JNK) protein. Results The relative expression of Calnexin mRNA: the experimental group, the empty vector group and the control group were 0.28, 0.01, 1.01, 0.06, 1.00 and 0.08 respectively; the experimental group was lower than that of the empty vector group and the control group (P0.05). The relative expression of GRP78, IRE1, TRAF2, ASK1, p-ASK1, JNK and p-JNK was higher than that in the control group (P <0.05). However, there was no statistical difference between the empty vector group and the control group (P <0.05). Conclusion Calnexin can induce endoplasmic reticulum stress and activate the apoptosis pathway of JNK cells, thus promoting the apoptosis of SK-NEP-1 cells.
【作者单位】：海南省人民医院;
【基金】：海南省自然科学基金资助项目(20168283)
【分类号】：R737.11

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