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山东大学学报（医学版） ›› 2016, Vol. 54 ›› Issue (2): 44-48.doi: 10.6040/j.issn.1671-7554.0.2015.077

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原癌基因c-jun对毒胡萝卜素内酯处理的小鼠胚胎成纤维细胞生存率的影响

张洁1,王爱红2,董波2,赵鹏2   

  1. 山东大学附属省立医院 1.营养科;2.心内科, 山东 济南 250021
  • 收稿日期:2015-04-24 出版日期:2016-02-10 发布日期:2016-02-10
  • 通讯作者: 赵鹏. E-mail:pengalfie@163.com E-mail:pengalfie@163.com
  • 基金资助:
    国家自然科学基金青年科学基金(81200176);山东省优秀中青年科学家科研奖励基金(BS2010YY046)

c-jun affects thapsigargin-induced mouse fibroblasts survival

ZHANG Jie1, WANG Aihong2, DONG Bo2, ZHAO Peng2   

  1. 1. Department of Nutrition;
    2. Department of Cardiology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China
  • Received:2015-04-24 Online:2016-02-10 Published:2016-02-10

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摘要： 目的 观察原癌基因c-jun对经毒胡萝卜素内酯(TG)处理的小鼠胚胎成纤维细胞生存率的影响,并探讨其机制。 方法 应用内质网应激诱导剂TG、钙调神经磷酸酶(CaN)的结构活性形式CnA(pCMV-CnA)和附加血凝素表位的c-jun表达载体(pSRα-HA-c-jun)处理基因敲除型(c-jun-/-)和基因重组型(c-jun Re)两组小鼠胚胎成纤维细胞后观察细胞生存率;蛋白免疫印迹技术分析蛋白c-jun、p-c-jun、JNK、p-JNK、Adapt78和α-tubulin的表达。 结果 经TG处理引起p-JNK与p-c-jun在c-jun Re细胞中的强烈激活,而CaN活性在c-jun-/-细胞中显著增加(P<0.05);CaN通过pCMV-CnA的外源性表达导致Adapt78表达在TG处理的c-jun Re细胞中有增强;在c-jun-/-细胞中由TG与pCMV- CnA同时处理后其细胞生存率低于同样处理后的c-jun Re细胞(P<0.05);c-jun外显基因在c-jun-/-细胞的过表达,同时引起了c-jun在靶细胞的磷酸化及诱导产生的Adapt78表达上调,c-jun基因的外部表达抑制了TG诱导的细胞死亡。 结论 c-jun基因表达水平的改变影响TG诱导的小鼠胚胎成纤维细胞生存率的变化,其变化与细胞内CaN和Adapt78蛋白表达的改变有关,c-jun基因表达增强导致Adapt78蛋白表达增强,从而减弱CaN活性,并产生出抵抗TG诱导细胞死亡的保护机制。

关键词: 钙调神经磷酸酶, 细胞生存率, c-jun基因, Adapt78蛋白

Abstract: Objective To examine the impact of c-jun on thapsigargin-induced mouse fibroblasts survival and its mechanism. Methods Thapsigargin, pCMV-CnA and pSRα-HA-c-jun were used to treat c-jun-/- and c-jun Re mouse fibroblasts. Cell viability was evaluated with trypan blue dye exclusion. Calcineurin assays were performed according to the manufacturers instructions. c-jun, p-c-jun, JNK, p-JNK, Adapt78 and α-tubulin were analyzed with immunoblotting. Results TG treatment led to strong JNK activation and phosphorylation of c-jun in c-jun Re cells. We observed a higher level of calcineurin activity in c-jun-/- cells compared with c-jun Re cells(P<0.05). TG treatment resulted in significantly increased calcineurin activity in c-jun-/- cells but not in c-jun Re cells. Exogenous expression of calcineurin via pCMV-CnA caused increased Adapt78 expression in both c-jun-/- and c-jun Re cells, although TG treatment induced Adapt78 expression only in c-jun Re cells. Adapt78 expression induced by pCMV-CnA in c-jun-/- cells was associated with protection against TG-induced cell death. Overexpression of exogenous c-jun in c-jun-/- cells caused both an autophosphorylation of c-jun in the target cells and exogenous expression of c-jun recovered thapsigargin-induced Adapt78 up-regulation and inhibited TG-induced cell death. Conclusion The change of c-jun gene expression level can affect TG-induced mouse cell survival rate. The changes of calcineurin and Adapt78 protein expression are related to the 山 东 大 学 学 报 (医 学 版)54卷2期 -张洁,等.原癌基因c-jun对毒胡萝卜素内酯处理的小鼠胚胎成纤维细胞生存率的影响 \=-alteration of c-jun gene expression. The increased c-jun gene expression results in enhanced expression of Adapt78 protein, thus weakens the calcineurin activity and produces protective mechanism against TG-induced cell death.

Key words: Calcineurin, Adapt78 protein, Cell survival rate, c-jun gene

中图分类号: 

  • R541
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