Passegue E, Laverriere J N, Gourdji D
Groupe de Biologie de la Cellule Neuroendocrine, CNRS URA 1115, Collège de France, Paris.
Mol Cell Neurosci. 1994 Apr;5(2):109-18. doi: 10.1006/mcne.1994.1013.
The hypothalamic neuropeptide TRH stimulates PRL release and PRL gene transcription in GH3B6 rat pituitary cells. In this model, TRH also raises c-fos proto-oncogene mRNA levels. c-fos is assumed to be involved in the transduction of external signals to the nucleus as a component of AP1 transcription factor, a complex that contains a member of the jun proto-oncogene family. Hence, the main aim of this study was to look for the expression of c-jun, jun B, and jun D proto-oncogenes in TRH-stimulated GH3B6 cells, compared to c-fos mRNA levels and PRL secretion. This was performed using serum-starved cells. Northern blotting and dot blotting revealed that jun B mRNA were expressed at noticeable levels in control cells and were strongly stimulated by TRH. This stimulation was dose-dependent and exhibited an early peak and a long-lasting plateau phase stabilized at 4 h, similar to TRH-induced c-fos mRNA induction. c-jun mRNA was expressed at minute levels in control cells and was transiently stimulated by TRH. The jun D message was not detected. TRH-induced jun B and c-fos responses were characteristic of immediate early genes as shown by the superinduction observed under cycloheximide treatment and the total inhibition elicited by actinomycin D. Finally, the responses to serum at 30 min and 4 h revealed differences among the proto-oncogenes studied and indicate that the action of TRH is specific.(ABSTRACT TRUNCATED AT 250 WORDS)
下丘脑神经肽TRH可刺激GH3B6大鼠垂体细胞释放催乳素(PRL)并促进PRL基因转录。在此模型中,TRH还可提高原癌基因c-fos的mRNA水平。c-fos被认为作为AP1转录因子的一个组分参与外部信号向细胞核的转导,AP1转录因子是一种包含jun原癌基因家族成员的复合体。因此,本研究的主要目的是,与c-fos mRNA水平和PRL分泌相比,探寻TRH刺激的GH3B6细胞中原癌基因c-jun、jun B和jun D的表达情况。这是使用血清饥饿细胞进行的。Northern印迹法和斑点印迹法显示,jun B mRNA在对照细胞中以显著水平表达,并受到TRH的强烈刺激。这种刺激呈剂量依赖性,呈现一个早期峰值和一个在4小时稳定的持久平台期,类似于TRH诱导的c-fos mRNA诱导。c-jun mRNA在对照细胞中表达水平极低,并受到TRH的短暂刺激。未检测到jun D信息。TRH诱导的jun B和c-fos反应具有即刻早期基因的特征,如在环己酰亚胺处理下观察到的超诱导以及放线菌素D引起的完全抑制所示。最后,在30分钟和4小时对血清的反应揭示了所研究原癌基因之间的差异,并表明TRH的作用具有特异性。(摘要截短至250字)