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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

Digestive Diseases and Sciences volume 58, pages 1016–1025 (2013)Cite this article

Abstract

Background

The 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.

Aims

Our purpose is to confirm the anti-invasion and anti-migration effects of the down-regulation of Notch1 in HCC cells.

Methods

The 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.

Results

In 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.

Conclusions

Down-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.

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Acknowledgments

We are grateful to Fuqin Zhang who provided me the technical help. This work was supported by grants from the National Natural Science Foundation of China (Grants No. 30872480) and the Major Program of the National Natural Science Foundation of China (Grants No. 81030010/H0318).

Conflict of interest

None.

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Affiliations

  1. Department of Hepatobiliary Surgery, Xijing Hospital, The Fourth Military Medical University, Xi’an, 710032, Shannxi, People’s Republic of China

    Liang Zhou, De-sheng Wang, Qing-jun Li, Wei Sun, Yong Zhang & Ke-feng Dou

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  1. Liang Zhou
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  2. De-sheng Wang
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  3. Qing-jun Li
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  4. Wei Sun
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Correspondence to Ke-feng Dou.

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Liang Zhou, De-sheng Wang and Qing-jun Li contributed equally to this work and should be recognized as co-first authors.

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Zhou, L., Wang, Ds., Li, Qj. et al. 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. Dig Dis Sci 58, 1016–1025 (2013). https://doi.org/10.1007/s10620-012-2434-7

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  • DOI: https://doi.org/10.1007/s10620-012-2434-7

Keywords

  • Notch1 signaling pathway
  • E-cadherin
  • Snail
  • Cyclooxygenase-2
  • Invasion
  • Migration