Identification of rat cell lines that preferentially express insulin-like growth factor binding proteins rlGFBP-1, 2, or 3.
Identification of rat cell lines that preferentially express insulin-like growth factor binding proteins rlGFBP-1, 2, or 3.
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优先表达胰岛素样生长因子结合蛋白 rlGFBP-1、2 或 3 的大鼠细胞系的鉴定。
作者：
Yvonne W.;A. L. Brown;C. Orlowski;D. E. Graham;L. Y. Tseng;J. A. Romanus;M. Rechler
胰岛素样生长因子(IGF)的生物利用度和作用是由它们与特定的IGF结合蛋白(IGFBP)决定的。已分离到编码三种相关IGFBP的互补DNA克隆：人IGFBP-1(hIGFBP-1)、人IGFBP-3(hIGFBP-3)和大鼠IGFBP-2(rIGFBP-2)。IGFBP-1和IGFBP-3在人血浆中的调节不同，提示它们具有不同的功能。为了研究不同IGFBPs调控的分子基础，我们鉴定了一组表达单一优势结合蛋白的大鼠细胞系，并建立了区分不同结合蛋白的检测策略。用rIGFBP-2和hIGFBP-1的抗体进行免疫沉淀和免疫印迹检测条件培养液中的蛋白质，并与rIGFBP-2和hIGFBP-1的cDNAs杂交。1)C6胶质细胞和B104神经母细胞瘤细胞表达约40kDa的糖基化结合蛋白，很可能代表成年大鼠血清中150 kDa IGF：结合蛋白复合体的结合亚基rIGFBP-3。C6和B104结合蛋白不与rIGFBP-2抗体反应，来自C6和B104细胞的RNA不与rIGFBP-2或hIGFBP-1的cDNA探针杂交。2)来自正常大鼠肝脏的BRL-3A、Clone 9和TRL 12-15细胞表达rIGFBP-2，这是一种30 kDa的非糖基化IGF结合蛋白，可被rIGFBP-2抗体识别，但不被hIGFBP-1抗体识别。来自这些细胞的RNA与rIGFBP-2cDNA探针杂交，但不与hIGFBP-1探针杂交。3)H35大鼠肝癌细胞表达30 kDa非糖基化IGFBP，推测为rIGFBP-1。它不与rIGFBP-2的抗体发生反应，但可被hIGFBP-1的多克隆和单抗识别。来自H35细胞的RNA与hIGFBP-1cDNA探针杂交，但不与rIGFBP-2探针杂交。通过H35细胞系rIGFBP-1的表达，使我们能够建立和验证这种蛋白的特定检测方法，使我们能够研究其在完整大鼠中的调节。鉴定一组表达特定IGFBP的大鼠细胞系有助于阐明IGFBP调控的分子机制。
The bioavailability and action of the insulin-like growth factors (IGFs) are determined by specific IGF-binding proteins (IGFBP) to which they are complexed. Complementary DNA clones have been isolated that encode three related IGFBPs: human IGFBP-1 (hIGFBP-1), human IGFBP-3 (hIGFBP-3), and rat IGFBP-2 (rIGFBP-2). IGFBP-1 and IGFBP-3 are regulated differently in human plasma, suggesting that they have different functions. In order to study the molecular basis of the regulation of the different IGFBPs, we have identified a panel of rat cell lines that express a single predominant binding protein and developed an assay strategy to distinguish the different binding proteins. Proteins in conditioned medium were examined by ligand blotting, and by immunoprecipitation and immunoblotting using antibodies to rIGFBP-2 and hIGFBP-1; RNAs were hybridized to cDNA probes for rIGFBP-2 and hIGFBP-1. 1) C6 glial cells and B104 neuroblastoma cells express an approximately 40 kilodalton (kDa) glycosylated binding protein that most likely represents rIGFBP-3, the binding subunit of the 150 kDa IGF: binding protein complex in adult rat serum. The C6 and B104 binding proteins do not react with antibodies to rIGFBP-2, and RNAs from C6 and B104 cells do not hybridize to cDNA probes for rIGFBP-2 or hIGFBP-1. 2) BRL-3A, Clone 9, and TRL 12-15 cell lines derived from normal rat liver express rIGFBP-2, a 30 kDa nonglycosylated IGF-binding protein that is recognized by antibodies to rIGFBP-2 but not by antibodies to hIGFBP-1. RNAs from these cells hybridize to a rIGFBP-2 cDNA probe, but not to a hIGFBP-1 probe. 3) H35 rat hepatoma cells express a 30 kDa nonglycosylated IGFBP that is presumptively identified as rIGFBP-1. It does not react with antibodies to rIGFBP-2, but is recognized by polyclonal and monoclonal antibodies to hIGFBP-1. RNA from H35 cells hybridizes to a hIGFBP-1 cDNA probe, but not to a rIGFBP-2 probe. Expression of rIGFBP-1 by the H35 cell line has enabled us to establish and validate specific assays for this protein that allow us to study its regulation in intact rats. Identification of a panel of rat cell lines expressing specific IGFBPs should be useful in elucidating the molecular mechanisms of IGFBP regulation.
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DOI：
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发表时间：
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影响因子：
--
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作者：
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DOI：
--
发表时间：
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期刊：
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影响因子：
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DOI：
10.1016/0006-291x(89)92254-7
发表时间：
1989
影响因子：
3.1
作者：
Ocrant,I;Pham,H;Oh,Y;Rosenfeld,RG
通讯作者：
Rosenfeld,RG
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DOI：
10.1210/mend-2-5-404
发表时间：
1988
期刊：
Molecular endocrinology (Baltimore, Md.)
影响因子：
--
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Powell,DR