Dermody W C, Rosen M A, Ananthaswamy R, Levy A G, Hixson C V, Aldenderfer P H, Nelson-Rees W A, Marangos P J, Swanson E C
J Clin Endocrinol Metab. 1981 Nov;53(5):970-7. doi: 10.1210/jcem-53-5-970.
We have developed three human cloned cell lines that produce immunoreactive human calcitonin (ihCT) and ACTH (iACTH) as well as exhibit characteristics of cultured neural cells. Clones HMS-41/I, -78/2, and -98/2 were developed from cell lines HeLa AV3, MBA 9812 (bronchogenic carcinoma), and SW 267 (pheochromocytoma), respectively. Karyological analysis of both the parent and the cloned cell lines confirmed the identity of HeLa AV3 and MBA 9812. When grown in serum-free media designed for culturing neural cells, the patterns of production for both ihCT and iACTH varied among the clones. The multiple patterns of hormone production suggest that the mechanisms involved in the biosynthesis, processing, and secretion of these hormones differ among the clones. The clones contain neuron-specific enolase and the putative neurotransmitters beta-alanine and gamma-amino butyric acid, and they respond to cAMP analogs by differentiating, as noted by the extension of neurites (except the HeLa-derived HMS-41/I). The iACTH extracted from cells and synthetic ACTH exhibited equivalent profiles upon isoelectric focusing. The forms of ihCT noted in cell extracts were similar to those observed in extracts of human tumor tissue. Our rabbit antiserum to hCT failed to detect ihCT in those cell extracts prepared for ACTH determination or in extracts of rat pituitaries, but it did detect CT in rat thyroids by both RIA and immunofluorescent procedures. We concluded that our antisera to hCT do not detect the precursor form of ACTH. The availability of these cloned cell lines provides model systems for examining the production of these peptide hormones and the concomitant expression of neural and endocrine characteristics.
我们已经培育出三种人类克隆细胞系，它们能产生免疫反应性人降钙素（ihCT）和促肾上腺皮质激素（iACTH），并且具有培养神经细胞的特性。克隆株HMS - 41/I、- 78/2和- 98/2分别从细胞系HeLa AV3、MBA 9812（支气管癌）和SW 267（嗜铬细胞瘤）培育而来。对亲代细胞系和克隆细胞系的核型分析证实了HeLa AV3和MBA 9812的一致性。当在专为培养神经细胞设计的无血清培养基中生长时，ihCT和iACTH的产生模式在各克隆株之间有所不同。激素产生的多种模式表明，这些激素在生物合成、加工和分泌过程中涉及的机制在各克隆株之间存在差异。这些克隆株含有神经元特异性烯醇化酶以及假定的神经递质β - 丙氨酸和γ - 氨基丁酸，并且它们对环磷酸腺苷类似物有反应，表现为神经突延长（HeLa来源的HMS - 41/I除外）。从细胞中提取的iACTH和合成的ACTH在等电聚焦时表现出相同的图谱。在细胞提取物中发现的ihCT形式与在人类肿瘤组织提取物中观察到的相似。我们针对hCT的兔抗血清在用于ACTH测定的细胞提取物或大鼠垂体提取物中未能检测到ihCT，但通过放射免疫分析和免疫荧光程序在大鼠甲状腺中检测到了CT。我们得出结论，我们针对hCT的抗血清不能检测到ACTH的前体形式。这些克隆细胞系的可用性为研究这些肽类激素的产生以及神经和内分泌特征的伴随表达提供了模型系统。
