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瑞士3T3成纤维细胞中α1(I)-胶原蛋白基因表达对细胞黏附的响应调控

Regulation of alpha 1 (I)-collagen gene expression in response to cell adhesion in Swiss 3T3 fibroblasts.

作者信息

Dhawan J, Farmer S R

机构信息

Department of Biochemistry, Boston University School of Medicine, Massachusetts 02118.

出版信息

J Biol Chem. 1990 Jun 5;265(16):9015-21.

PMID:2188970
Abstract

Nonadhesive conditions cause Swiss 3T3 fibroblasts to enter a quiescent state that is reversed upon reattachment to a surface. Previously, we demonstrated that adhesion in serum-free conditions is sufficient to activate suspension-arrested cells out of Go, with the induction of the growth-associated genes, c-fos, c-myc, and actin. In this study, we have employed this system to identify programs of gene expression that respond primarily to the adhesive state of the cell, rather than the growth state. We show that cells in different adhesive states can be distinguished by their patterns of protein synthesis. Analysis of one adhesion-responsive protein led to its identification as pro-alpha 1 (I)-collagen. Pro-alpha 1 (I)-collagen synthesis and mRNA levels are decreased up to 6-fold in suspension-arrested fibroblasts, but are enhanced up to 5-fold as cells approach confluence. This suggests that the reduced expression in suspension-arrested cells is not simply a result of quiescence. In addition, reattachment of suspended cells in serum-free conditions caused a 7-fold induction of collagen mRNA levels and a greater than 20-fold rise in the rate of procollagen synthesis. The expression of c-myc was induced during adhesion in serum-free medium as well as by serum addition to suspension-arrested cells. However, alpha 1 (I)-collagen gene expression was unaffected by serum in the absence of adhesion. These results indicate that collagen gene expression is directly responsive to cell adhesion, independent of the growth state.

摘要

非黏附条件会使瑞士3T3成纤维细胞进入静止状态,而重新附着于表面后该状态会逆转。此前,我们证明在无血清条件下的黏附足以激活处于G0期的悬浮停滞细胞,诱导生长相关基因c-fos、c-myc和肌动蛋白的表达。在本研究中,我们利用该系统来鉴定主要对细胞黏附状态而非生长状态作出反应的基因表达程序。我们发现处于不同黏附状态的细胞可通过其蛋白质合成模式加以区分。对一种黏附反应蛋白的分析表明其为前α1(I)型胶原蛋白。悬浮停滞的成纤维细胞中前α1(I)型胶原蛋白的合成及mRNA水平降低多达6倍,但随着细胞接近汇合状态,其水平可增强多达5倍。这表明悬浮停滞细胞中表达的降低并非仅仅是静止状态的结果。此外,无血清条件下悬浮细胞的重新附着导致胶原蛋白mRNA水平诱导7倍,前胶原合成速率升高超过20倍。在无血清培养基中黏附过程中以及向悬浮停滞细胞中添加血清时,c-myc的表达均被诱导。然而,在无黏附的情况下,α1(I)型胶原蛋白基因表达不受血清影响。这些结果表明胶原蛋白基因表达直接对细胞黏附作出反应,与生长状态无关。

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