Eckes B, Mauch C, Hüppe G, Krieg T
Department of Dermatology, University of Cologne, Germany.
FEBS Lett. 1993 Mar 1;318(2):129-33. doi: 10.1016/0014-5793(93)80006-g.
Culturing human fibroblasts in a three-dimensional collagen matrix leads to a reduction of collagen I by more than 90%, both on the level of mRNA steady-state as well as protein. In order to differentiate changes in de novo transcription and posttranscriptional control, nuclear run on assays and pulse/chase experiments determining mRNA stability were used. Our results indicate that de novo transcription of the COL1A1 gene and pro-alpha 1 (I)collagen mRNA half-life are both decreased by 50% in fibroblasts grown in three-dimensional collagen lattices as compared to monolayer cultures. The extracellular matrix therefore elicits signals which are transduced from the cell surface to the inside of fibroblasts resulting in a specific reprogramming of transcriptional as well as posttranscriptional processes.
在三维胶原基质中培养人成纤维细胞会导致Ⅰ型胶原在mRNA稳态水平和蛋白质水平上均减少90%以上。为了区分从头转录和转录后调控的变化,采用了核转录分析以及测定mRNA稳定性的脉冲/追踪实验。我们的结果表明,与单层培养相比,在三维胶原晶格中生长的成纤维细胞中,COL1A1基因的从头转录和前α1(Ⅰ)胶原mRNA半衰期均降低了50%。因此,细胞外基质引发从细胞表面传导至成纤维细胞内部的信号,导致转录和转录后过程的特定重编程。