The Down-Regulation of Notch1 Inhibits the Invasion and Migration of Hepatocellular Carcinoma Cells by Inactivating the Cyclooxygenase-2/Snail/E-cadherin Pathway In Vitro
The Down-Regulation of Notch1 Inhibits the Invasion and Migration of Hepatocellular Carcinoma Cells by Inactivating the Cyclooxygenase-2/Snail/E-cadherin Pathway In Vitro
复制标题
体外下调Notch1通过灭活Cyclooxygenase-2/Snail/E-cadherin通路抑制肝细胞癌细胞的侵袭和迁移
DOI：
10.1007/s10620-012-2434-7
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发表时间：
2013-04-01
影响因子：
3.1
通讯作者：
Dou, Ke-feng
中科院分区：
文献类型：
--
作者：
Zhou, Liang;Wang, De-sheng;Dou, Ke-feng
背景Notch信号通路在肿瘤中起重要作用，但Notch1参与肝癌细胞侵袭和迁移的机制尚不清楚。目的证实Notch1下调在肝癌细胞中的抗侵袭和抗迁移作用。用RT-PCR和/或Western blotting检测Notch1、Notch1胞内区(N1ICD)、E-钙粘蛋白、Snail和COX-2的表达。RNA干扰下调NOTCH1和Snail的表达，NS-398抑制COX-2的表达。结果在肝癌细胞中，Snail、Notch1和COX-2表达上调，E-cadherin在mRNA和/或蛋白水平表达下调。Snail或Notch1的下调或COX-2的抑制可以增加E-钙粘蛋白的mRNA和蛋白的表达，降低肝癌细胞的侵袭和迁移能力。下调Notch1或抑制COX-2可降低Snail的mRNA和蛋白表达。Notch1的下调也可以减少COX-2的蛋白表达。然而，外源PGE2可以逆转下调的Notch1的作用。结论Notch1下调可能是通过调节COX-2使Snail/E-cadherin失活的有效途径，从而抑制肝癌细胞的侵袭和迁移。下调的Notch1对细胞侵袭和迁移的抑制作用不依赖于细胞凋亡。
BackgroundThe Notch signaling pathway plays an important role in cancer, but the mechanism by which Notch1 participates in invasion and migration of hepatocellular carcinoma (HCC) cells is unclear.AimsOur purpose is to confirm the anti-invasion and anti-migration effects of the down-regulation of Notch1 in HCC cells.MethodsThe invasion and migration capacities of HCC cells were detected with Transwell cell culture chambers. The expressions of Notch1, Notch1 intracellular domain (N1ICD), E-cadherin, Snail, and cyclooxygenase-2 (COX-2) were analyzed by RT-PCR and/or western blotting. Notch1 and Snail were down-regulated by RNA interference, and COX-2 was inhibited by NS-398. Cell apoptosis was analyzed by MTT and flow cytometry.ResultsIn HCC cells, Snail, Notch1, and COX-2 were up-regulated, and E-cadherin was down-regulated in mRNA and/or protein levels. The down-regulation of Snail or Notch1 or the inhibition of COX-2, respectively, can increase the mRNA and protein expressions of E-cadherin and decrease the invasion and migration capabilities of HCC cell. Down-regulated Notch1 or inhibited COX-2 can reduce the mRNA and protein expressions of Snail. The down-regulation of Notch1 can also reduce the protein expression of COX-2. However, exogenous PGE2 can reverse the role of down-regulated Notch1. The results of MTT and flow cytometry showed that down-regulated Notch1 did not affect HCC cell viability.ConclusionsDown-regulated Notch1 may be an effective approach to inactivating Snail/E-cadherin by regulating COX-2, which results in inhibiting the invasion and migration of HCC cells. The inhibitory effects of down-regulated Notch1 on cell invasion and migration were independent of apoptosis.