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化学转化大鼠成纤维细胞中的细胞内钙离子及钙离子结合蛋白

Intracellular Ca2+ and Ca(2+)-binding proteins in chemically transformed rat fibroblasts.

作者信息

Klug M, Blum J K, Ye Q, Berchtold M W

机构信息

Institute of Veterinary Biochemistry, University of Zürich, Switzerland.

出版信息

Exp Cell Res. 1994 Aug;213(2):313-8. doi: 10.1006/excr.1994.1204.

Abstract

Disturbed Ca2+ handling and altered levels of Ca(2+)-binding proteins (CaBPs) have been found in many transformed cells. To investigate whether there exists a correlation between expression of CaBPs and intracellular Ca2+ we analyzed three transformed rat fibroblast cell lines (T14, T10, and T43) exhibiting different levels of CaBPs. All three cell lines express calmodulin (CaM) at elevated levels compared to normal tissues. In addition, oncomodulin (OM) is expressed at high levels in T14 cells and at much lower levels in T10 cells, while T43 cells do not express OM. We demonstrate that CaBP levels are affected by changes of extracellular [Ca2+] ([Ca2+]o). In reduced [Ca2+]o, CaM levels were increased up to 60%, while OM levels were decreased up to 65% in T14 and T10 cells. Increase of [Ca2+]o, on the other hand, led to a decrease of CaM levels and an increase of OM levels, suggesting that [Ca2+]o exerts its effect on OM and CaM differently. Comparison of the growth rates in reduced [Ca2+]o revealed that T14 cells with the most pronounced CaM increase induced by low [Ca2+]o grew best under these conditions. These results indicate that elevated CaM expression but not the presence of OM reduces the Ca2+ requirement for growth. Intracellular [Ca2+] transients are not buffered by the high concentration of CaBPs in T14 cells. In contrast, [Ca2+]i transients induced by increase of [Ca2+]o or addition of serum were most pronounced in T14 cells compared to T10 and T43 cells.

摘要

在许多转化细胞中都发现了钙处理紊乱和钙结合蛋白(CaBP)水平改变的情况。为了研究CaBP的表达与细胞内钙离子之间是否存在相关性,我们分析了三种表现出不同CaBP水平的转化大鼠成纤维细胞系(T14、T10和T43)。与正常组织相比,这三种细胞系均高表达钙调蛋白(CaM)。此外,癌调蛋白(OM)在T14细胞中高表达,在T10细胞中表达水平低得多,而T43细胞不表达OM。我们证明CaBP水平受细胞外钙离子浓度([Ca2+]o)变化的影响。在降低的[Ca2+]o条件下,T14和T10细胞中的CaM水平增加高达60%,而OM水平降低高达65%。另一方面,[Ca2+]o的增加导致CaM水平降低和OM水平增加,这表明[Ca2+]o对OM和CaM的作用方式不同。在降低的[Ca2+]o条件下比较生长速率发现,在低[Ca2+]o诱导下CaM增加最明显的T14细胞在这些条件下生长最好。这些结果表明,CaM表达升高而非OM的存在降低了生长所需的钙离子浓度。T14细胞中高浓度的CaBP并未缓冲细胞内钙离子瞬变。相反,与T10和T43细胞相比,在T14细胞中,由[Ca2+]o增加或添加血清诱导的[Ca2+]i瞬变最为明显。

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