Garcia-Montero A, Vasseur S, Mallo G V, Soubeyran P, Dagorn J C, Iovanna J L
Centre de Recherche INSERM, EMI 0116, Marseille/France.
Eur J Cell Biol. 2001 Nov;80(11):720-5. doi: 10.1078/0171-9335-00209.
We report here that the mere fact of changing culture medium for fresh medium induced in several cell lines the expression of stress-activated genes including protein kinases p38, JNK and ERK1/2 and the transcription factor C/EBPbeta. As a consequence, p8, a gene induced by stress in several tissues, was strongly up-regulated. Induction did not occur after change for cell-conditioned medium. Induction was however transient, with a peak at 60 min for p38, at 15-30 min for JNK and at 15 min for ERK1/2, at 2-3 hours for C/EBPbeta and at 4-6 hours for p8. Repression of the induction was due to the secretion of thermolabile molecule(s) that progressively conditioned the medium. As low as 25% of conditioned medium added to fresh culture medium was sufficient to abolish the stress response. Taken together, our data indicate that the renewal of culture medium induces a transient cellular stress that may be a source of artifacts in experiments performed shortly after a change of culture medium.
我们在此报告,仅仅是将培养基换成新鲜培养基这一行为,就在几种细胞系中诱导了应激激活基因的表达,这些基因包括蛋白激酶p38、JNK和ERK1/2以及转录因子C/EBPβ。结果,p8(一种在多种组织中由应激诱导的基因)被强烈上调。换成细胞条件培养基后未出现诱导现象。然而,诱导是短暂的,p38在60分钟时达到峰值,JNK在15 - 30分钟时达到峰值,ERK1/2在15分钟时达到峰值,C/EBPβ在2 - 3小时时达到峰值,p8在4 - 6小时时达到峰值。诱导的抑制是由于分泌了逐渐调理培养基的热不稳定分子。向新鲜培养基中添加低至25%的条件培养基就足以消除应激反应。综上所述,我们的数据表明,培养基的更新会诱导短暂的细胞应激,这可能是在更换培养基后不久进行的实验中出现假象的一个来源。