TRANSFORMATION OF HUMAN CILIARY EPITHELIAL-CELLS BY SIMIAN VIRUS-40 - INDUCTION OF CELL-PROLIFERATION AND RETENTION OF BETA-2-ADRENERGIC RECEPTORS
TRANSFORMATION OF HUMAN CILIARY EPITHELIAL-CELLS BY SIMIAN VIRUS-40 - INDUCTION OF CELL-PROLIFERATION AND RETENTION OF BETA-2-ADRENERGIC RECEPTORS
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DOI：
10.1073/pnas.83.22.8754
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发表时间：
1986-11-01
影响因子：
11.1
通讯作者：
WAX, MB
中科院分区：
文献类型：
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作者：
COCAPRADOS, M;WAX, MB
人眼睫状体上皮细胞经猴病毒40转化后，经多代传代培养成功。细胞克隆8-SVHCE被分离出来，并通过免疫沉淀和药理学研究证明在该组织的亲本细胞中存在几种功能特性。免疫沉淀显示大肿瘤(T)抗原的存在，Southern印迹分析显示病毒DNA被掺入高分子量睫状体上皮细胞DNA。放射性标记的拮抗剂[125I]碘吲哚与8-SVHCE细胞的膜制剂(Kd=41.8 pm，Bmax=67.1 fmol/mg蛋白质)直接结合，证明了β-肾上腺素能受体的存在。与[125I]碘吲哚和选择性药物的竞争实验表明，受体是β2-肾上腺素能亚型。8-SVHCE中儿茶酚胺刺激细胞产生cAMP和异丙肾上腺素依赖的蛋白磷酸化Vimentin的研究表明，β-肾上腺素能受体介导的过程在体内睫状体上皮调节房水产生方面具有重要的功能转换。
Ciliary epithelial cells derived from human eye were successfully propagated through many generations after transformation with simian virus 40. The cell clone 8-SVHCE was isolated and characterized by immunoprecipitation and pharmacological studies that demonstrated the presence of several functional properties observed in the parent cells of this tissue. Immunoprecipitation revealed the presence of large tumor (T) antigen, and Southern blot analysis showed the incorporation of viral DNA into high molecular weight ciliary epithelial cell DNA. The presence of .beta.-adrenergic receptors was demonstrated by direct binding of a radiolabeled antagonist, [125I]iodopindolol, to membrane preparations of 8-SVHCE cells (Kd = 41.8 pM and Bmax = 67.1 fmol/mg of protein). Competition experiments with [125I]iodopindolol and selective drugs suggested that the receptors are of the .beta.2-adrenergic subtype. Studies of catecholamine-stimulated cellular cAMP production and of isoproterenol-dependent protein phosphorylation of vimentin in 8-SVHCE indicated the functional conversation of .beta.-adrenergic receptor-mediated processes that are thought to be important in the regulation of aqueous humor production by the ciliary epithelium in vivo.