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Issue 12, 2016
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Oxygen as a chemoattractant: confirming cellular hypoxia in paper-based invasion assays

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Abstract

Low oxygen tension, or hypoxia, is a common occurrence in solid tumors. Hypoxia is a master regulator of cellular phenotype, and is associated with increased tumor invasion and aggressiveness as well as adverse patient prognosis. Oxygen has recently been linked with the selective movement of different cancer cell types in three-dimensional invasion assays utilizing paper-based scaffolds. It has remained unclear, however, if cells in these paper-based invasion assays are experiencing hypoxia. In this manuscript, we adapted cell-based methods to measure oxygen tension in our 3D invasion assays: the adduction of pimonidazole to free thiols in the cell, indicative of a reducing environment; the localization of hypoxia inducible factors to the nucleus; and the expression of hypoxia-regulated gene products. We utilized each method to compare the oxygen tension in different locations of the paper-based invasion stacks and found an oxygen gradient is indeed forming. Specifically, we found that the extent of pimonidazole binding, as well as the levels and activities of nucleus-localized HIF-α proteins, increase as the distance between the cells and the source of fresh medium increases. These complementary cell-based readouts not only confirm the selective invasion we observe is due to an oxygen gradient, they also show the gradient is temporal in nature and evolves with increasing culture period.

Graphical abstract: Oxygen as a chemoattractant: confirming cellular hypoxia in paper-based invasion assays

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Supplementary files

Article information


Submitted
16 Mar 2016
Accepted
25 Apr 2016
First published
26 Apr 2016

Analyst, 2016,141, 3874-3882
Article type
Paper

Oxygen as a chemoattractant: confirming cellular hypoxia in paper-based invasion assays

A. S. Truong and M. R. Lockett, Analyst, 2016, 141, 3874
DOI: 10.1039/C6AN00630B

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