Tumor promoter 12-O-tetradecanoylphorbol-13-acetate receptors in normal human transitional epithelial cells.
Tumor promoter 12-O-tetradecanoylphorbol-13-acetate receptors in normal human transitional epithelial cells.
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正常人移行上皮细胞中的肿瘤启动子 12-O-十四烷酰佛波醇-13-乙酸酯受体。
DOI：
10.1093/carcin/6.3.427
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发表时间：
1985
期刊：
影响因子：
4.7
通讯作者：
Reznikoff,CA
中科院分区：
文献类型：
--
作者：
Verma,AK;Bryan,GT;Reznikoff,CA
为了研究TPA对培养的人尿路上皮细胞（HUC）体外转化的促进作用，我们对HUC原代培养的肿瘤启动子TPA受体进行了表征。[3H]TPA与完整活HUC特异性结合；在37°c下，在约30分钟内达到最大特异性结合。[3H]TPA以饱和和竞争的方式与HUC结合。与完整细胞特异性结合的Scatchard分析显示单个斜率对应于平衡解离常数（Kd）为0.56 nM；饱和tpa结合容量为2.37 pmol/106HUC （1.43 × 106个位点/细胞）。[3H]TPA与HUC的颗粒组分具有特异性和高亲和力；装订是可饱和和可逆的。[3H]TPA的特异性结合在4°C ~ 1 nM处发生饱和。与颗粒组分特异性结合的Scatchard分析显示单一斜率对应的Kd为1.08 nM；饱和时tpa结合能力为2.05 pmol/mg蛋白（75万分子/ HUC）。[3H]TPA的结合被具有生物活性的phorbol酯（phorbol didecanoate）抑制，而无活性的phorbol不竞争TPA的结合。糖精钠、表皮生长因子、视黄酸和地塞米松均不影响结合。[3H]TPA特异性地与HUC细胞质部分结合，但仅在钙和磷脂酰丝氨酸存在的情况下。在HUC馏分中检测到钙活化和磷脂敏感的蛋白激酶活性。这些结果表明在HUC中存在高亲和力的TPA特异性受体。
As a prelude to study the promotion with TPA ofin vitrotransformation of human urothelial cells (HUC) in culture, we characterized tumor promoter TPA receptors in primary cultures of HUC. [3H]TPA bound specifically to intact living HUC; maximum specific binding was attained in ∼30 min at 37°C. [3H]TPA bound to HUC in a saturable and competitive manner. Scatchard analysis of specific binding to intact cells displayed a single slope corresponding to an equilibrium dissociation constant (Kd) of 0.56 nM; at saturation TPA-binding capacity was 2.37 pmol/106HUC (1.43 × 106sites per cell). [3H]TPA bound specifically and with high affinity to the particulate fractions of HUC; binding was both saturable and reversible. Saturation of the specific binding of [3H]TPA occurred at ∼1 nM at 4°C. Scatchard analysis of specific binding to the particulate fraction displayed a single slope corresponding to a Kd of 1.08 nM; at saturation TPA-binding capacity was 2.05 pmol/mg protein (750 000 molecules per HUC). [3H]TPA binding was inhibited by the biologically active phorbol ester, phorbol didecanoate, whereas inactive phorbol did not compete for TPA binding. Binding was not affected by sodium saccharin, epidermal growth factor, retinoic acid or dexamethasone. [3H]TPA bound specifically to the HUC cytosolic fraction but only in the presence of calcium and phosphatidylserine. Calcium-activated and phospholipid-sensitive protein kinase activity was detected in HUC fractions. These results indicate the presence of high-affinity specific receptors for TPA in HUC.