PI3-K/Akt/JNIUNF-κB is essential for MMP-9 expression and outgrowth in human limbal epithelial cells on intact amniotic membrane
PI3-K/Akt/JNIUNF-κB is essential for MMP-9 expression and outgrowth in human limbal epithelial cells on intact amniotic membrane
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DOI：
10.1016/j.scr.2012.02.005
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发表时间：
2012-07-01
影响因子：
1.2
通讯作者：
Yang, Chuen-Mao
中科院分区：
文献类型：
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作者：
Cheng, Ching-Yi;Hsieh, Hsi-Lung;Yang, Chuen-Mao
基质金属蛋白酶-9（MMP-9）在完整羊膜（AM）上扩增的人鼓膜上皮细胞生长过程中起重要作用。MMP-9的表达和细胞生长的机制仍不清楚。在这里，我们证明，MMP-9是优先表达在前沿的鼓膜上皮生长。用PI 3-K（LY 294002）、Akt（SH-5）、MEK 1/2（U 0126）和JNK 1/2（SP 600125）的抑制剂处理减弱了生长区域，表明PI 3-K/Akt、p42/p44 MAPK和JNK 1/2参与了完整AM扩增的鼓膜上皮细胞的生长。然而，MMP-9在转录和翻译水平上的表达被SP 600125、LY 294002或SH-5处理减弱，而不是被U 0126和SB 202190处理，这表明JNK 1/2和PI 3-K/Akt参与MMP-9的表达。此外，NF-κ B磷酸化和核转位在前缘特别明显，这通过用SP 600125或LY 294002处理而减弱。选择性NF-κ B抑制剂Helenalin可降低鼓膜上皮生长和MMP-9表达。最后，数据显示PI 3-K/Akt是MMP-9表达中JNK 1/2通路的上游组分。因此，在该模型中，MAPK和PI 3-K/Akt都是完整AM上的鼓膜上皮生长所必需的，只有PI 3-K/Akt/JNK是通过激活转录因子NF-κ B介导的MMP-9表达所必需的。(C)2012 Elsevier B. V.保留所有权利。
Matrix metalloproteinase-9 (MMP-9) plays an important role in the outgrowth of expanded human timbal epithelial cells on intact amniotic membranes (AM). The mechanisms of MMP-9 expression and cell outgrowth remain unknown. Here, we demonstrated that MMP-9 is preferentially expressed at the leading edge of timbal epithelial outgrowth. Treatment with the inhibitors of PI3-K (LY294002), Akt (SH-5), MEK1/2 (U0126), and JNK1/2 (SP600125) attenuated the outgrowth area, indicating that PI3-K/Akt, p42/p44 MAPK, and JNK1/2 are involved in the outgrowth of intact AM-expanded timbal epithelial cells. However, MMP-9 expression at both transcriptional and translational levels was attenuated by treatment with SP600125, LY294002, or SH-5, not by U0126 and SB202190, suggesting that JNK1/2 and PI3-K/Akt participate in MMP-9 expression. Moreover, NF-kappa B phosphorylation and nuclear translocation was especially noted at the leading edge, which was attenuated by treatment with SP600125 or LY294002. Helenalin, a selective NF-kappa B inhibitor, reduced both the timbal epithelial outgrowth and MMP-9 expression. Finally, the data reveal that PI3-K/Akt is an upstream component of the JNK1/2 pathway in MMP-9 expression. Thus, both MAPKs and PI3-K/Akt are required for timbal epithelial outgrowth on intact AM, only the PI3-K/Akt/JNK is essential for MMP-9 expression mediated through activation of transcriptional factor NF-kappa B in this model. (C) 2012 Elsevier B.V. All rights reserved.