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通过与玻璃珠和琼脂糖珠共价结合的药物和激素对培养的肿瘤细胞进行亲和分离。

Affinity isolation of cultured tumor cells by means of drugs and hormones covalently bound to glass and Sepharose beads.

作者信息

Venter B R, Venter J C, Kaplan N O

出版信息

Proc Natl Acad Sci U S A. 1976 Jun;73(6):2013-7. doi: 10.1073/pnas.73.6.2013.

DOI:10.1073/pnas.73.6.2013
PMID:180534
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC430438/
Abstract

Isoproterenol, corticotropin (ACTH), and triodothyronine immobilized on glass and Sepharose beads by diazotization procedures have been shown to interact with cultured tumor cells of "target tissue" origin. Cells used were rat glioma cells (C6), rat adrenal tumor cells (Y-1), and rat pituitary tumor cells (GH3). The rat glioma cells bound principally to immobilized isoproterenol, whereas the rat adrenal tumor cells bound to immobilized corticotropin, and rat pituitary tumor cells bound to immobilized triiodothyronine. Binding was inhibited by preincubation of the cells in soluble drug or hormone. With C6 cells there was a positive correlation between adenylate cyclase [ATP pyrophosphate-lyase (cyclizing, EC 4.6.1.1] stimulation and the degree of binding to the immobilized isoproterenol. Norepinephrine, bound through the ethanolamine side chain via an amide linkage, did not bind cells, demonstrating specific structural requirements for drug-cell interactions. HeLa cells were shown to bind tightly to diphtheria toxin coupled to Sepharose beads via an amide bond. This binding was inhibited by prior incubation of the Sepharose toxin with purified antitoxin. Toxin bound to Sepharose via an azo bond did not bind cells. These data suggest that the cell affinities are due to cell surface receptors interacting with the immobilized drugs and hormones, and that the observed affinities possibly reflect the relative receptor complement of these cells.

摘要

通过重氮化程序固定在玻璃珠和琼脂糖珠上的异丙肾上腺素、促肾上腺皮质激素（ACTH）和三碘甲状腺原氨酸已被证明能与源自“靶组织”的培养肿瘤细胞相互作用。所使用的细胞为大鼠胶质瘤细胞（C6）、大鼠肾上腺肿瘤细胞（Y-1）和大鼠垂体肿瘤细胞（GH3）。大鼠胶质瘤细胞主要与固定化的异丙肾上腺素结合，而大鼠肾上腺肿瘤细胞与固定化的促肾上腺皮质激素结合，大鼠垂体肿瘤细胞与固定化的三碘甲状腺原氨酸结合。细胞在可溶性药物或激素中预孵育可抑制结合。对于C6细胞，腺苷酸环化酶[ATP焦磷酸裂解酶（环化，EC 4.6.1.1）]的刺激与对固定化异丙肾上腺素的结合程度之间存在正相关。通过乙醇胺侧链经酰胺键结合的去甲肾上腺素不与细胞结合，这表明药物与细胞相互作用存在特定的结构要求。已证明HeLa细胞能紧密结合通过酰胺键与琼脂糖珠偶联的白喉毒素。这种结合可通过用纯化的抗毒素预先孵育琼脂糖毒素来抑制。通过偶氮键与琼脂糖结合的毒素不与细胞结合。这些数据表明细胞亲和力是由于细胞表面受体与固定化药物和激素相互作用所致，并且观察到的亲和力可能反映了这些细胞的相对受体组成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/ef6fba2612c3/pnas00036-0249-f.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/5f2bbf985083/pnas00036-0249-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/c3112bb6cbd1/pnas00036-0249-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/1a17f79c28de/pnas00036-0249-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/02839ccb97ea/pnas00036-0249-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/2a48d8b163f9/pnas00036-0249-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/ef6fba2612c3/pnas00036-0249-f.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/5f2bbf985083/pnas00036-0249-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/c3112bb6cbd1/pnas00036-0249-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/1a17f79c28de/pnas00036-0249-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/02839ccb97ea/pnas00036-0249-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/2a48d8b163f9/pnas00036-0249-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9297/430438/ef6fba2612c3/pnas00036-0249-f.jpg

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本文引用的文献

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Binding of estrogen receptor to estradiol immobilized on insoluble resins.雌激素受体与固定在不溶性树脂上的雌二醇的结合。
Biochim Biophys Acta. 1969 Apr 29;176(3):626-31. doi: 10.1016/0005-2760(69)90229-x.
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Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads.通过亲和色谱法进行蛋白质纯化。琼脂糖和聚丙烯酰胺珠粒的衍生化。
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Separation of specific antibody-forming mouse cells by their adherence to insolubilized endogenous hormones.通过将特异性抗体形成小鼠细胞与固定化内源性激素结合来进行分离。
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8
Affinity binding of intact fat cells and their ghosts to immobilized insulin.完整脂肪细胞及其细胞膜空壳与固定化胰岛素的亲和结合。
Proc Natl Acad Sci U S A. 1973 Mar;70(3):792-6. doi: 10.1073/pnas.70.3.792.
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Biologically active catecholamines covalentyly bound to glass beads.与玻璃珠共价结合的生物活性儿茶酚胺。
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10
Site of action of adrenocorticotropic hormone (ACTH) in adrenal cell cultures.促肾上腺皮质激素（ACTH）在肾上腺细胞培养物中的作用位点。
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