Doxycycline Alters Metabolism and Proliferation of Human Cell Lines

Ethan Ahler,Heather R. Christofk,Autumn G. York,Ashley Cass,Steven J. Bensinger,Daniel Braas,Thomas G. Graeber,William J. Sullivan,Rajeev Samant

doi:10.1371/journal.pone.0064561

Abstract

The tetracycline antibiotics are widely used in biomedical research as mediators of inducible gene expression systems. Despite many known effects of tetracyclines on mammalian cells–including inhibition of the mitochondrial ribosome–there have been few reports on potential off-target effects at concentrations commonly used in inducible systems. Here, we report that in human cell lines, commonly used concentrations of doxycycline change gene expression patterns and concomitantly shift metabolism towards a more glycolytic phenotype, evidenced by increased lactate secretion and reduced oxygen consumption. We also show that these concentrations are sufficient to slow proliferation. These findings suggest that researchers using doxycycline in inducible expression systems should design appropriate controls to account for potential confounding effects of the drug on cellular metabolism.

Highlights

The tetracycline family is a class of broad-spectrum antibiotics that have been used clinically since the mid-twentieth century
Because the tetracyclines have been shown to inhibit matrix metalloproteinases, retard proliferation, induce apoptosis, and impair mitochondrial function in various experimental settings, we were interested to determine whether these drugs can alter cellular metabolism at concentrations commonly used in inducible systems [4,5,6,7,8,9,10,11,12]
Despite a reportedly weak interaction between the antibiotics and the mitochondrial ribosome, at high concentrations they have been shown to impair synthesis of proteins encoded in the mitochondrial genome–many of which are involved in oxidative metabolism–and promote a shift towards glycolysis [4]

Summary

Introduction

The tetracycline family is a class of broad-spectrum antibiotics that have been used clinically since the mid-twentieth century. Since they have found application beyond their antimicrobial activity in both the clinic and biomedical research [1,2,3]. They have found application beyond their antimicrobial activity in both the clinic and biomedical research [1,2,3] They are widely used in the latter context as mediators of inducible gene expression systems, but often with little discussion of or control for potential off-target effects they may have on mammalian cells. Despite a reportedly weak interaction between the antibiotics and the mitochondrial ribosome, at high concentrations they have been shown to impair synthesis of proteins encoded in the mitochondrial genome–many of which are involved in oxidative metabolism–and promote a shift towards glycolysis [4]

Methods

Results

Conclusion