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Nitric oxide mediates cell aggregation and mesenchymal to epithelial transition in anoikis-resistant lung cancer cells

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Abstract

Cancer cell aggregation has been long known to facilitate metastatic potential of cancer cells. In addition, the presence of nitric oxide (NO) in cancer area may have a significant impact on aggregation behavior of the cells. We show herein that lung cancer H460 cells possessing high ability of anoikis resistance formed loose aggregates in detached condition. Importantly, NO treatment tightened the aggregates by enhancing cell–cell interaction via E-cadherin-dependent mechanism, and such E-cadherin contact increased anoikis resistance potential by up-regulating pro-survival signals of the cells including active ATP-dependent tyrosine kinase and extracellular-regulated protein kinases (ERK1/2). Since an increase of E-cadherin was frequently found in mesenchymal to epithelial transition (MET) process, we further tested the cells for MET markers and found that NO treatment of these cells significantly enhanced MET. As aggregation and MET of cancer cells may facilitate cancer metastasis by many means, the insights gained from the present study could benefit the deep understanding in the biology of cancer cell metastasis.

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Abbreviations

AKT:

ATP-dependent tyrosine kinase

Cav-1:

Caveolin-1

NO:

Nitric oxide

XTT:

2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt

E-cad:

E-cadherin

N-cad:

N-cadherin

Vim:

Vimentin

p-AKT:

Phosphorylated AKT

PBS:

Phosphate-buffered saline

Mcl-1:

Myeloid cell leukemia sequence 1

TBST:

Tris-buffered saline with 0.1 % Tween

PI:

Propidium iodide

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Acknowledgments

The authors would like to thank the 100th Anniversary Chulalongkorn University Fund for Doctoral Scholarship, Ratchadaphiseksompot Endowment Fund, Chulalongkorn University (RES560530132-HR), and Mr. Krich Rajprasit, proofreader.

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Authors and Affiliations

  1. Department of Pharmacology and Physiology and Cell-based Drug and Health Product Development Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand

    Phattrakorn Powan

  2. Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand

    Pithi Chanvorachote

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  1. Phattrakorn Powan
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Correspondence to Pithi Chanvorachote.

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Powan, P., Chanvorachote, P. Nitric oxide mediates cell aggregation and mesenchymal to epithelial transition in anoikis-resistant lung cancer cells. Mol Cell Biochem 393, 237–245 (2014). https://doi.org/10.1007/s11010-014-2066-7

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  • DOI: https://doi.org/10.1007/s11010-014-2066-7

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