In vitro induction of human skin ornithine decarboxylase by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate.
In vitro induction of human skin ornithine decarboxylase by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate.
复制标题
肿瘤启动子 12-O-十四烷酰佛波醇-13-乙酸酯体外诱导人皮肤鸟氨酸脱羧酶。
DOI：
10.1093/jnci/75.1.85
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
P. Carbone
中科院分区：
文献类型：
--
作者：
A. Verma;C. Loprinzi;R. Boutwell;P. Carbone
建立了12-O-十四酰佛波醇-13-乙酸酯（TPA）体外诱导人皮肤穿刺活检标本中鸟氨酸脱羧酶（ODC）活性的方法。将TPA添加到含有从手术标本获得的单个3-mm人皮肤穿刺活检样品的1 ml无血清最低必需培养基中导致ODC活性的诱导，在TPA添加后5小时具有峰值活性。体外诱导人表皮ODC活性依赖于培养基中TPA的浓度;加入0.1 μ M TPA后6小时观察到ODC活性增加约2倍，加入1 μ M TPA观察到ODC活性增加约5倍。TPA还导致来自健康志愿者的3 mm皮肤穿刺活检样品中ODC活性增加约5倍至6倍。人皮肤穿刺活检样品即使在4 ° C下在无血清培养基中储存24小时也对TPA诱导的ODC活性保持响应。无论是完整的普通人表皮还是可溶性表皮提取物用于ODC测定，都观察到TPA对ODC活性的类似程度的诱导。ODC活性增加是酶活性ODC蛋白增加的结果，通过[3 H]二氟甲基鸟氨酸结合试验定量。因此，人类皮肤，像小鼠皮肤一样，对用于ODC诱导的TPA有响应。
A method was developed for the in vitro induction by 12-O-tetradecanoylphorbol-13-acetate (TPA) of ornithine decarboxylase (ODC) activity in human skin punch biopsy samples. Addition of TPA to 1 ml serum-free minimum essential medium containing a single 3-mm human skin punch biopsy sample obtained from a surgical specimen resulted in an induction of ODC activity with a peak activity at 5 hours after TPA addition. In vitro induction of human epidermal ODC activity was dependent on the TPA concentration in the medium; about a twofold increase in ODC activity was observed 6 hours after the addition of 0.1 microM TPA, and about a fivefold increase in ODC activity was observed with 1 microM TPA. TPA also caused about a fivefold to sixfold increase in ODC activity in 3-mm skin punch biopsy samples from healthy volunteers. Human skin punch biopsy samples remained responsive to TPA induction of ODC activity even when stored in serum-free medium at 4 degrees C for 24 hours. A similar degree of induction of ODC activity by TPA was observed whether whole unfractionated human epidermis or a soluble epidermal extract was used for ODC assays. Increased ODC activity was the result of the increase in enzymatically active ODC protein, quantitated by a [3H]difluoromethylornithine-binding assay. Thus human skin, like mouse skin, is responsive to TPA for ODC induction.