Filvaroff E, Stern D F, Dotto G P
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510.
Mol Cell Biol. 1990 Mar;10(3):1164-73. doi: 10.1128/mcb.10.3.1164-1173.1990.
Very little is known about early molecular events triggering epithelial cell differentiation. We have examined the possible role of tyrosine phosphorylation in this process, as observed in cultures of primary mouse keratinocytes after exposure to calcium or 12-O-tetradecanoylphorbol-13-acetate (TPA). Immunoblotting with phosphotyrosine-specific antibodies as well as direct phosphoamino acid analysis revealed that induction of tyrosine phosphorylation occurs as a very early and specific event in keratinocyte differentiation. Very little or no induction of tyrosine phosphorylation was observed in a keratinocyte cell line resistant to the differentiating effects of calcium. Treatment of cells with tyrosine kinase inhibitors prevented induction of tyrosine phosphorylation by calcium and TPA and interfered with the differentiative effects of these agents. These results suggest that specific activation of tyrosine kinase(s) may play an important regulatory role in keratinocyte differentiation.
关于触发上皮细胞分化的早期分子事件,人们了解甚少。我们研究了酪氨酸磷酸化在这一过程中可能发挥的作用,这一作用在原代小鼠角质形成细胞暴露于钙或12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)后的培养物中有所观察。使用磷酸酪氨酸特异性抗体进行免疫印迹以及直接的磷酸氨基酸分析表明,酪氨酸磷酸化的诱导是角质形成细胞分化过程中非常早期且特异性的事件。在对钙的分化作用具有抗性的角质形成细胞系中,几乎未观察到酪氨酸磷酸化的诱导,或根本没有诱导。用酪氨酸激酶抑制剂处理细胞可阻止钙和TPA诱导的酪氨酸磷酸化,并干扰这些试剂的分化作用。这些结果表明,酪氨酸激酶的特异性激活可能在角质形成细胞分化中起重要的调节作用。