Effects of cell surface receptor-altering agents on the binding and biological activity of 12-O-tetradecanoylphorbol-13-acetate in isolated epidermal cells.
Effects of cell surface receptor-altering agents on the binding and biological activity of 12-O-tetradecanoylphorbol-13-acetate in isolated epidermal cells.
复制标题
细胞表面受体改变剂对分离表皮细胞中 12-O-十四烷酰佛波醇-13-乙酸酯的结合和生物活性的影响。
DOI：
10.1093/carcin/3.10.1149
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发表时间：
1982
期刊：
影响因子：
4.7
通讯作者：
Boutwell,RK
中科院分区：
文献类型：
--
作者：
Perchellet,JP;Kishore,GS;Conrad,EA;Boutwell,RK
将分离的表皮细胞与已知干扰或改变细胞表面受体的超微结构的多种化合物一起孵育，并研究了12-O-十四酰基-佛波醇-13-乙酸酯（TPA）结合这些细胞并诱导表皮鸟氨酸脱羧酶（ODC）活性的能力。α-和β-肾上腺素能拮抗剂酚妥拉明和普萘洛尔以及胆碱能拮抗剂阿托品有效竞争[3 H]二氢-α-麦角隐亭、[3 H]二氢烯丙洛尔和[14 C]乙酰胆碱的结合受体，但不抑制TPA对ODC活性的诱导或[3 H]TPA与细胞的特异性结合。神经氨酸酶处理引起唾液酸从细胞中的时间和剂量相关性释放，并增强霍乱毒素对基础和TPA诱导的ODC活性的刺激作用，与单唾液酸神经节苷脂GM 1一样多。神经氨酸酶和其他膜改变剂，岩藻糖苷酶，半乳糖苷酶，半乳糖氧化酶，磷脂酶A2和C，和NaIO 4，单独使用和/或在我们的研究中的各种组合。所有测试的治疗抑制several 125 I标记的激素和表皮生长因子的细胞的特异性结合。相反，在相同的细胞系统中，这些处理都不能影响TPA的结合或生物活性。因此，这些结果表明，TPA在质膜水平上的主要相互作用以及其在完整细胞中的生物学效应不通过肾上腺素能或胆碱能受体进行，并且不需要细胞表面糖缀合物和磷脂的完整性。此外，视黄酸对TPA诱导的ODC活性的抑制作用不受上述处理的影响，表明视黄酸添加不太可能在质膜水平上干扰TPA相互作用。
Isolated epidermal cells were incubated with a variety of compounds known to interfere with or alter the ultrastructure of cell surface receptors, and the ability of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) to bind to these cells and induce epidermal ornithine decarboxylase (ODC) activity was investigated. The α- and β-adrenergic antagonists, phentolamine and propranolol, and the cholinergic antagonist, atropine, which competed effectively for the binding receptors of [3H]dihydro-α-ergocryptine, [3H]dihydroalprenolol, and [14C]acetylcholine, did not inhibit the induction of ODC activity by TPA or the specific binding of [3H]TPA to the cells. Neuraminidase treatments caused a time- and dose- related release of sialic acid from the cells and enhanced the stimulatory effect of cholera toxin on basal and TPA-induced ODC activities as much as the monosialoganglioside GM1. Neuraminidase and the other membrane-altering agents, fucosidase, galactosidase, galactose oxidase, phospholipases A2 and C, and NaIO4, were used alone and/or in various combinations in our studies. All treatments tested inhibited the specific binding of several125I-labeled hormones and epidermal growth factor to the cells. In contrast, none of these treatments was able, in the same cell system, to affect either the binding or the biological activity of TPA. Therefore, these results suggest that the primary interaction of TPA at the plasma membrane level as well as its biological effect in the intact cell do not proceed through adrenergic or cholinergic receptors and do not require the integrity of the cell surface glycoconjugates and phospholipids. In addition, the inhibitory effect of retinoic acid on TPA-induced ODC activity remained unaffected by some of the above treatments, suggesting that retinoic add is unlikely to interfere with TPA interactions at the plasma membrane level.