Powell M B, Rosenberg R K, Graham M J, Birch M L, Yamanishi D T, Buckmeier J A, Meyskens F L
Arizona Cancer Center, University of Arizona, Tucson 85724.
J Cancer Res Clin Oncol. 1993;119(4):199-206. doi: 10.1007/BF01624431.
Normal human melanocytes require 12-O-tetradecanoylphorbol 13-acetate (TPA) for prolonged growth in vitro. In contrast, the growth of human malignant melanoma cells is often inhibited by TPA. In this study, we have confirmed and extended these observations. Since protein kinase C (PKC) is an important mediator of the effects of TPA, we have investigated the nature of this differential growth response by examining PKC expression and activity in primary cultures of human neonatal melanocytes and metastatic melanoma cell strains. PKC, when measured by immunoreactivity or a functional assay, was found to be more abundant in melanoma cells than in melanocytes. When specific isotypes were examined by Northern analysis, PKC-alpha and -epsilon were expressed in both melanocytes and melanoma. PKC-beta was expressed in melanocytes, but was undetectable by Northern analysis in 10 out of 11 melanoma cell strains. Southern analysis revealed that no gross deletions or rearrangements of the PKC-beta gene had occurred. These data suggest that down-regulation of the PKC-beta gene occurs frequently during the process of transformation of melanocytes. Furthermore, differential expression of PKC isotypes may explain the different effects of TPA on melanocyte and melanoma cell growth.
正常人类黑素细胞在体外长期生长需要12 - O -十四烷酰佛波醇-13 -乙酸酯(TPA)。相比之下,人恶性黑色素瘤细胞的生长常被TPA抑制。在本研究中,我们证实并扩展了这些观察结果。由于蛋白激酶C(PKC)是TPA作用的重要介质,我们通过检测人新生儿黑素细胞原代培养物和转移性黑色素瘤细胞系中的PKC表达及活性,研究了这种生长差异反应的本质。通过免疫反应性或功能测定法测量时,发现PKC在黑色素瘤细胞中比在黑素细胞中更为丰富。当通过Northern分析检测特定亚型时,PKC -α和 -ε在黑素细胞和黑色素瘤中均有表达。PKC -β在黑素细胞中表达,但在11个黑色素瘤细胞系中的10个中,Northern分析未检测到其表达。Southern分析显示PKC -β基因未发生明显缺失或重排。这些数据表明,在黑素细胞转化过程中,PKC -β基因经常发生下调。此外,PKC亚型的差异表达可能解释了TPA对黑素细胞和黑色素瘤细胞生长的不同影响。