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Acid treatment of melanoma cells selects for invasive phenotypes

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Abstract

Solid tumors become acidic due to hypoxia and upregulated glycolysis. We have hypothesized that this acidosis leads to more aggressive invasive behavior during carcinogenesis (Nature Reviews Cancer 4:891–899, 2004). Previous work on this subject has shown mixed results. While some have observed an induction of metastasis and invasion with acid treatments, others have not. To investigate this, human melanoma cells were acclimated to low pH growth conditions. Significant cell mortality occurred during acclimation, suggesting that acidosis selected for resistant phenotypes. Cells maintained under acidic conditions exhibited a greater range of motility, a reduced capacity to form flank tumors in SCID mice and did not invade more rapidly in vitro, compared to non-selected control cells. However, re-acclimation of these selected cells to physiological pH gave rise to stable populations with significantly higher in vitro invasion. These re-acclimated cells maintained higher invasion and higher motility for multiple generations. Transcriptomic analyses of these three phenotypes revealed significant differences, including upregulation of relevant pathways important for tissue remodeling, cell cycle control and proliferation. These results reinforce the hypothesis that acidosis promotes selection of stable, more invasive phenotypes, rather than inductive changes, which would be reversible.

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Abbreviations

DMEM:

Dulbecco’s Modified Eagle’s Medium

MAPK:

Mitogen-activated protein kinase

MMP:

Matrix metalloproteinase

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Acknowledgements

NIH R01 CA077575 (RJG); NIH R01 CA093650 (RAG); Beckman Foundation Undergraduate Fellowship (KB); Howard Hughes Medical Institute grant #52003749 (REM).

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Author notes

  1. Raymond E. Moellering

    Present address: Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA, 02138, USA

Authors and Affiliations

  1. Arizona Cancer Center, Arizona Health Sciences Center, Tucson, USA

    Raymond E. Moellering, Kvar C. Black, Chetan Krishnamurty, Brenda K. Baggett & Robert J. Gillies

  2. Translational Genomics Research Institute (TGen), 445 N. 5th St, Phoenix, AZ, 85004, USA

    Phillip Stafford

  3. Agilent Technologies, 5301 Stevens Creek Blvd, Santa Clara, CA, 95051, USA

    Matthew Rain

  4. Department of Radiology, Arizona Cancer Center, 1515 N Campbell Ave, Tucson, AZ, 85724-5024, USA

    Robert A. Gatenby & Robert J. Gillies

Authors
  1. Raymond E. Moellering
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  2. Kvar C. Black
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  8. Robert J. Gillies
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Corresponding author

Correspondence to Robert J. Gillies.

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10585_2008_9145_MOESM1_ESM.tif

Supplemental Figure 1. Quantitative fluorescent tracking of invasion. MB-MDA-231 cells were acid-selected and tested for invasive potential exactly as discussed in Materials and Methods for C8161 melanoma cells and Figure 4. (TIF 2043 kb)

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Moellering, R.E., Black, K.C., Krishnamurty, C. et al. Acid treatment of melanoma cells selects for invasive phenotypes. Clin Exp Metastasis 25, 411–425 (2008). https://doi.org/10.1007/s10585-008-9145-7

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  • DOI: https://doi.org/10.1007/s10585-008-9145-7

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