MITOGENIC EFFECTS OF THYROTROPIN AND ADENOSINE-3',5'-MONOPHOSPHATE IN DIFFERENTIATED NORMAL HUMAN THYROID-CELLS INVITRO
MITOGENIC EFFECTS OF THYROTROPIN AND ADENOSINE-3',5'-MONOPHOSPHATE IN DIFFERENTIATED NORMAL HUMAN THYROID-CELLS INVITRO
复制标题
DOI：
10.1210/jcem-66-6-1158
复制
发表时间：
1988-06-01
影响因子：
5.8
通讯作者：
DUMONT, JE
中科院分区：
文献类型：
--
作者：
ROGER, P;TATON, M;DUMONT, JE
以往对培养的人甲状腺细胞（主要来自病理组织）的研究未能证明TSH的有丝分裂作用，因此提出TSH在体内的生长作用可能是间接的。为了重新研究TSH对DNA合成和细胞增殖的影响，我们建立了9例正常甲状腺组织的原代培养。在添加1%血清的培养基中，通过胶原酶/疾病消化释放的甲状腺滤泡发育为单层细胞，通过聚集和细胞质收缩对TSH作出反应。当在无血清培养基中播种时，细胞仍以密集的聚集体存在，周围有少数缓慢扩散的细胞。在后一种情况下，细胞通过显示非常高的碘捕获水平，对TSH和其他cAMP生成刺激物（如霍乱毒素和福斯克林）做出反应。暴露于血清中不可逆转地消除了这种分化功能。TSH刺激1%血清培养细胞增殖（倍增时间由106 h减少至76 h），使[3H]胸苷标记指数提高100%（每培养皿DNA含量）。在无血清培养的细胞中（最初用血清播种后的密集聚集体或细胞单层），对照DNA合成水平较低，根据胸苷苷掺入酸沉淀物质的测量和放射自显影上标记细胞核的计数（24小时内标记细胞核的数量高达40%），100 mU/L TSH可产生高达8倍的DNA合成刺激（20 mU/L TSH也能检测到刺激）。TSH的有丝分裂作用需要高胰岛素浓度（8.3倍）。10-7 mol/L)或低胰岛素样生长因子I浓度。TSH的有丝分裂作用部分被霍乱毒素、福斯可林和二丁基cAMP所模仿。表皮生长因子和肉豆酸酯也能刺激甲状腺细胞增殖和DNA合成，但能有效抑制tsh刺激的碘离子转运。我们得出结论，TSH至少部分通过cAMP起作用，是人类甲状腺细胞的一种有效生长因子，因此为TSH在体内形成甲状腺肿的作用提供了体外实验基础。
Previous studies of human thyroid cells in culture (mostly from pathological tissues) failed to demonstrate mitogenic effect of TSH, leading to the proposal that the growth effect of TSH in vivo might be indirect. To reexamine the influence of TSH on DNA synthesis and cell proliferation, we established primary cultures of normal thyroid tissue from nine subjects. When seeded in a 1% serum-supplemented medium, thyroid follicles released by collagenase/dispase digestion developed as a cell monolayer that responded to TSH by rounding up and by cytoplasmic retraction. When seeded in serum-free medium, the cells remained associated in dense aggregates surrounded by few slowly spreading cells. In the latter condition, the cells responded to TSH and other stimulators of cAMP production, such as cholera toxin and forskolin, by displaying very high iodide-trapping levels. Exposure to serum irreversibly abolished this differentiated function. TSH stimulated the proliferation (as shown by DNA content per culture dish) of 1% serum cultured cells (doubling times were reduced from 106 to 76 h) and increased by 100% the [3H]thymidine labeling indices. In serum-free cultured cells (dense aggregates or cell monolayers after initial seeding with serum), contral levels of DNA synthesis were lower, and up to 8-fold stimulation of DNA synthesis occurred in response to 100 mU/L TSH (stimulation was consistently detected with 20 mU/L), based on measurements of [3H]thymidine incorporation into acid-precipitable material and counts of labeled nuclei on autoradiographs (up to 40% labeled nuclei within 24 h). The mitogenic effect of TSH required a high insulin concentration (8.3 .times. 10-7 mol/L) or a low insulin-like growth factor I concentration. The mitogenic effects of TSH were mimicked in part by cholera toxin, forskolin, and dibutyryl cAMP. Epidermal growth factor and phorbol myristate ester also stimulated thyroid cell proliferation and DNA synthesis, but they potently inhibited TSH-stimulated iodide transport. We conclude that TSH, acting at least in part through cAMP, is a potent growth factor for human thyroid cells and thus provide an experimental basis in vitro for the well established in vivo goitrogenic action of TSH.