Subclones of a rat parathyroid cell line (PT‐r): Regulation of growth and production of parathyroid hormone‐related peptide (PTHRP)
Subclones of a rat parathyroid cell line (PT‐r): Regulation of growth and production of parathyroid hormone‐related peptide (PTHRP)
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大鼠甲状旁腺细胞系 (PT–r) 的亚克隆：甲状旁腺激素相关肽 (PTHRP) 生长和产生的调节
DOI：
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发表时间：
1990
影响因子：
6.2
通讯作者：
M. Brandi
中科院分区：
文献类型：
--
作者：
K. Sakaguchi;K. Ikeda;Francesco Curcio;G. Aurbach;M. Brandi
从大鼠甲状旁腺细胞系（PT-r细胞）中分离出4个亚克隆，并检查了形态和功能特征。亚克隆1和2显示多边形形状，显示对钙的生长和分泌反应（分别在1.2和1.7 mM下的半最大供应量），并对促胰液素产生cAMP产生（比基础高14.5倍和16.9倍）和激素分泌（比基础高41%和58%）的反应。亚克隆4是细长的形式，不响应钙或胰泌素。亚克隆3显示混合形态、细长和多边形形状，对钙（1.7 mM时的半数最大抑制）和胰泌素（cAMP，3.2倍增加和激素分泌，比基础增加50%）有中度反应。检测克隆中代表甲状旁腺激素（PTH）和甲状旁腺激素相关肽（PTHRP）的信使RNA（mRNA）含量。仅发现PTHRPmRNA，释放的肽几乎都是PTHRP。即使用敏感的RNA探针也未检测到PTH mRNA。PTHRP的mRNA的量与从每个克隆释放到培养基中的PTH样生物活性的量密切相关（在含有0.7 mM离子钙的培养基中，144.7 ± 12.1、110.0 ± 12.9、68.0 ± 5.6和39.9 ± 2.4 pgEq的大鼠PTH-（-34）/107个细胞/12 h，来自亚克隆1、2、3和4，分别）。培养条件，低密度传代（小于1：50分流比）或高密度传代（大于1：10分流比），影响克隆1和2的形态和功能。在高密度培养3个月后，它们变得像亚克隆4一样伸长和功能去分化。去分化后，形态和功能特性不可逆的高钙，1，25-（OH）2D 3，或视黄酸。这些克隆可能有助于研究表型表达、钙调节、基因表达以及这些功能之间的相互关系。
Four subclones from a rat parathyroid cell line (PT‐r cell) have been isolated, and morphological and functional characteristics have been examined. Subclones 1 and 2 display a polygonal shape, show growth and secretory responses to calcium (half‐maximal suppressions at 1.2 and 1.7 mM, respectively), and respond to secretin with cAMP production (14.5‐fold and 16.9‐fold over basal) and hormone secretion (41 and 58% over basal). Subclone 4 is elongated in form and does not respond to calcium or secretin. Subclone 3 shows mixed morphology, elongated and polygonal shapes, with moderate response to calcium (half‐maximal suppression at 1.7 mM) and secretin (cAMP, 3.2‐fold increase and hormone secretion, 50% increase over basal). The clones were tested for content of messenger RNA (mRNA) representing parathyroid hormone (PTH) and parathyroid hormone‐related peptide (PTHRP). Only PTHRP mRNA was found. The peptide released is virtually all PTHRP. PTH mRNA was not detected even with a sensitive RNA probe. The amount of mRNA for PTHRP closely paralleled the amount of PTH‐like bioactivity released into the medium from each clone (144.7 ± 12.1, 110.0 ± 12.9, 68.0 ± 5.6, and 39.9 ± 2.4 pgEq of rat PTH‐(‐34) per 107 cells per 12 h in a medium with 0.7 mM ionized calcium, from subclones 1, 2, 3, and 4, respectively). Culture conditions, low‐density passage (less than 1:50 split ratio) or high‐density passage (greater than 1:10 split ratio), affected morphology and function of the clones 1 and 2. They became elongated and functionally dedifferentiated like subclone 4 after 3 months of high‐density culture. Upon dedifferentiation, morphological and functional properties were not reversible with high calcium, 1,25‐(OH)2D3, or retinoic acid. These clones may be useful for studies of phenotypic expression, calcium regulation, and gene expression and interrelationships among these functions.
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DOI：
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发表时间：
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期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
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DOI：
10.1016/0006-291x(89)91969-4
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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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影响因子：
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