Protein kinase C isozymes regulate proliferation and high cell density-mediated differentiation in HaCaT keratinocytes
Protein kinase C isozymes regulate proliferation and high cell density-mediated differentiation in HaCaT keratinocytes
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DOI：
10.1111/j.0906-6705.2003.00097.x
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发表时间：
2003-12-01
影响因子：
3.6
通讯作者：
Bíró, T
中科院分区：
文献类型：
--
作者：
Papp, H;Czifra, G;Bíró, T
蛋白激酶C（PKC）亚型在正常人表皮角质形成细胞（NHEK）分化的调节中起着关键作用。在这项研究中，我们调查的参与PKC系统的增殖和高细胞密度诱导分化的人永生化角质形成细胞系HaCaT。HaCaT角质形成细胞具有特征性的PKC亚型模式（PKCalpha、beta、gamma、delta、beta、eta、theta、zeta），其在增殖和分化期间改变。选择性PKC抑制剂GF 109203 X化合物抑制晚期（颗粒细胞）分化标志物外皮蛋白（INV）和聚丝蛋白（FIL）以及终末标志物角质细胞特异性转氨酶-1（TG）的表达，但不影响早期（棘细胞）标志物角蛋白10（K10）的水平和细胞增殖。PKC激活剂佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）可抑制细胞增殖，升高细胞内钙离子浓度，降低K10的表达，增加INV、FIL和TG的表达。这些数据表明，PKC的内源性激活调节晚期分化标志物的表达，并且PMA对PKC的外源性激活导致终末分化的诱导。由于PMA的细胞效应伴随着敏感的PKC亚型在增殖和分化培养中的差异下调，我们的研究结果认为，现有的PKC亚型在调节细胞增殖和高细胞密度诱导的HaCaT细胞分化中的差异作用。
Protein kinase C (PKC) isoforms play pivotal roles in the regulation of differentiation of normal human epidermal keratinocytes (NHEK). In this study, we investigated the participation of the PKC system in the proliferation and high cell density-induced differentiation of the human immortalized keratinocyte line HaCaT. HaCaT keratinocytes possessed a characteristic PKC isoform pattern (PKCalpha, beta, gamma,delta,epsilon,eta,theta,zeta), which altered during proliferation and differentiation. The GF109203X compound, a selective PKC inhibitor, suppressed the expressions of the late (granular cell) differentiation markers involucrin (INV) and filaggrin (FIL), and the terminal marker keratinocytes-specific transglutaminase-1 (TG), but did not affect the level of the early (spinous cell) marker keratin 10 (K10) and cellular proliferation. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, inhibited proliferation, elevated intracellular calcium concentration, decreased the expression of K10, and increased the expressions of INV, FIL, and TG. These data indicate that the endogenous activation of PKC regulates the expressions of the late differentiation markers, and that the exogenous activation of PKC by PMA results in the induction of terminal differentiation. Because the cellular effects of PMA were accompanied by differential down-regulations of the sensitive PKC isoforms in proliferating and differentiating cultures, our findings argue for the differential roles of the existing PKC isoforms in the regulation of cellular proliferation and high cell density-induced differentiation of HaCaT cells.