Ando Y, Lazarus G S, Jensen P J
Department of Dermatology, University of Pennsylvania School of Medicine, Philadelphia 19104-6142.
J Cell Physiol. 1993 Sep;156(3):487-96. doi: 10.1002/jcp.1041560308.
The involvement of protein kinase C (PKC) in epidermal growth factor (EGF)-induced human keratinocyte migration was studied with the phagokinetic assay. It was concluded that PKC activation does not mediate, but rather inhibits, EGF-induced keratinocyte migration. The following experimental observations support these conclusions: 1) The PKC inhibitor H-7 did not inhibit EGF-induced migration but instead led to a modest enhancement. 2) PKC activators such as phorbol-12-myristate-13-acetate (PMA), phorbol-12,13-dibutyrate (PDBu), and 1,2-dioctanoly-sn-glycerol inhibited migration, but biologically inactive 4 alpha-PMA had no effect. 3) PMA did not inhibit keratinocyte attachment and spreading but blocked migration almost immediately after addition. 4) Migration of PKC-depleted cells, which were produced by prolonged treatment with PDBu, was enhanced similarly to normal cells by EGF. 5) PKC-depleted cells were not susceptible to the inhibitory effects of phorbol esters on migration. Additional experiments, in which cells were preactivated with EGF, suggested that PKC inhibits the EGF effect at a post-receptor level. The inhibitory effect of PKC on keratinocyte migration was not restricted to EGF-induced migration; PKC activation also inhibited keratinocyte migration induced by bovine pituitary extract, insulin, insulin-like growth factor-1, and keratinocyte growth factor.
利用吞噬运动测定法研究了蛋白激酶C（PKC）在表皮生长因子（EGF）诱导的人角质形成细胞迁移中的作用。得出的结论是，PKC激活并不介导而是抑制EGF诱导的角质形成细胞迁移。以下实验观察结果支持这些结论：1）PKC抑制剂H-7不抑制EGF诱导的迁移，反而导致适度增强。2）PKC激活剂如佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）、佛波醇-12,13-二丁酸酯（PDBu）和1,2-二辛酰-sn-甘油抑制迁移，但无生物学活性的4α-PMA没有作用。3）PMA不抑制角质形成细胞的附着和铺展，但在添加后几乎立即阻止迁移。4）用PDBu长期处理产生PKC缺失的细胞，其迁移与正常细胞一样被EGF增强。5）PKC缺失的细胞对佛波醇酯对迁移的抑制作用不敏感。另外的实验中，细胞先用EGF预激活，结果表明PKC在受体后水平抑制EGF的作用。PKC对角质形成细胞迁移的抑制作用不限于EGF诱导的迁移；PKC激活也抑制牛垂体提取物、胰岛素、胰岛素样生长因子-1和角质形成细胞生长因子诱导的角质形成细胞迁移。
