MITOGENIC, MELANOGENIC, AND CAMP RESPONSES OF CULTURED NEONATAL HUMAN MELANOCYTES TO COMMONLY USED MITOGENS
MITOGENIC, MELANOGENIC, AND CAMP RESPONSES OF CULTURED NEONATAL HUMAN MELANOCYTES TO COMMONLY USED MITOGENS
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DOI：
10.1002/jcp.1041500226
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发表时间：
1992-02-01
影响因子：
5.6
通讯作者：
NORDLUND, JJ
中科院分区：
文献类型：
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作者：
ABDELMALEK, Z;SWOPE, VB;NORDLUND, JJ
已进行以下研究以比较和关联12-0-十四酰基佛波醇乙酸酯（TPA）、碱性成纤维细胞生长因子（bFGF）、霍乱毒素（CT）和异丁基甲基黄嘌呤（IBMX）对新生人类黑素细胞（NHM）增殖、酪氨酸酶活性和环磷酸腺苷（cAMP）浓度的影响。 NHM在含有8 nM TPA、200 ng/ml CT和10（-4）M IBMX的培养基中以最大速率增殖。 TPA单独没有导致最佳的黑素细胞增殖，并且如前所述，其促有丝分裂作用通过单独或同时添加CT和IBMX而大大增强。 人重组bFGF可替代TPA在NHM培养基中生长。 使用3 ng/ml hrbFGF、20 ng/ml CT和10（-4）M IBMX实现最大增殖。 1.2 ng/ml hrbFGF的促有丝分裂作用在CT和IBMX的伴随存在下而非单独存在下增强。 TPA单独在CT和IBMX的情况下引起酪氨酸酶活性的剂量依赖性刺激。 在0.8 nM TPA、20 ng/ml CT和10（-4）M IBMX存在下获得最大酪氨酸酶活性。 与TPA不同，单独使用hrbFGF会抑制酪氨酸酶活性。 在hrbFGF的存在下，酪氨酸酶活性增强CT和IBMX，但不是由CT单独。 TPA和hrbFGF单独使用均不能增加细胞内cAMP水平。 TPA比hrbFGF更能增强CT和IBMX对细胞内cAMP浓度的影响。 在我们的实验条件下，在hrbFGF的存在下，CT而不是IBMX导致cAMP浓度的剂量依赖性增加。 对NHM的进一步研究将旨在确定蛋白激酶C（PKC）在调节增殖和黑素生成中的确切作用以及hrbFGF激活的机制。
The following studies have been undertaken to compare and correlate the effects of 12-0-tetradecanoylphorbol acetate (TPA), basic fibroblast growth factor (bFGF), cholera toxin (CT), and isobutyl methylxanthine (IBMX) on neonatal human melanocyte (NHM) proliferation, tyrosinase activity, and cyclic adenosine monophosphate (cAMP) concentration. NHM proliferated at a maximal rate in medium containing 8 nM TPA, 200 ng/ml CT, and 10(-4) M IBMX. TPA alone did not result in optimal melanocyte proliferation, and, as previously shown, its mitogenic effect was greatly enhanced by the addition of CT and IBMX individually or concomitantly. Human recombinant (hr) bFGF could replace TPA in the NHM growth medium. Maximal proliferation was achieved using 3 ng/ml hrbFGF, 20 ng/ml CT, and 10(-4) M IBMX. The mitogenic effect of 1.2 ng/ml hrbFGF was potentiated in the concomitant but not individual presence of CT and IBMX. TPA alone in the absence of CT and IBMX caused a dose-dependent stimulation of tyrosinase activity. Maximal tyrosinase activity was obtained in the presence of 0.8 nM TPA, 20 ng/ml CT, and 10(-4) M IBMX. Unlike TPA, hrbFGF alone resulted in inhibition of tyrosinase activity. In the presence of hrbFGF, tyrosinase activity was potentiated by CT and IBMX, but not by CT alone. Neither TPA nor hrbFGF alone could increase intracellular cAMP levels. The effects of CT and IBMX on intracellular cAMP concentration were enhanced to a greater extent by TPA than by hrbFGF. Under our experimental conditions, in the presence of hrbFGF, CT but not IBMX resulted in a dose-dependent increase in cAMP concentration. Further studies on NHM will be aimed at determining the exact role of protein kinase C (PKC) in regulating proliferation and melanogenesis and the mechanism(s) activated by hrbFGF.