Doxycycline Impairs Mitochondrial Function and Protects Human Glioma Cells from Hypoxia-Induced Cell Death: Implications of Using Tet-Inducible Systems

International Journal of Molecular Sciences
Anna-Luisa LugerMichael W Ronellenfitsch

Abstract

Inducible gene expression is an important tool in molecular biology research to study protein function. Most frequently, the antibiotic doxycycline is used for regulation of so-called tetracycline (Tet)-inducible systems. In contrast to stable gene overexpression, these systems allow investigation of acute and reversible effects of cellular protein induction. Recent reports have already called for caution when using Tet-inducible systems as the employed antibiotics can disturb mitochondrial function and alter cellular metabolism by interfering with mitochondrial translation. Reprogramming of energy metabolism has lately been recognized as an important emerging hallmark of cancer and is a central focus of cancer research. Therefore, the scope of this study was to systematically analyze dose-dependent metabolic effects of doxycycline on a panel of glioma cell lines with concomitant monitoring of gene expression from Tet-inducible systems. We report that doxycycline doses commonly used with inducible expression systems (0.01⁻1 µg/mL) substantially alter cellular metabolism: Mitochondrial protein synthesis was inhibited accompanied by reduced oxygen and increased glucose consumption. Furthermore, doxycycline protected human glioma ...Continue Reading

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Citations

Mar 27, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Anton MisakKarol Ondrias
Jun 9, 2020·The Journal of Comparative Neurology·Min Ji HanJeong Tae Do
Mar 24, 2020·British Journal of Cancer·Anna L EngelAnna-Luisa Luger
Aug 31, 2020·Critical Reviews in Biotechnology·Aarti DoshiPatrick C Cirino
Oct 6, 2020·Frontiers in Physiology·Zhibo GaiMichele Visentin
Nov 26, 2020·Open Biology·Bernice A AganaDaniel R Schoenberg
Apr 4, 2021·International Journal of Molecular Sciences·Sonia Romero-CorderoConstanza Morén
Apr 4, 2021·Pathogens·Magdalena Dunowska, Sayani Ghosh
Aug 13, 2021·Molecular Biology of the Cell·Jolan De Boeck, Catherine Verfaillie

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Methods Mentioned

BETA
FCS
xenografts
transfections
transfection
Reverse Transcription-PCR
PCR
flow cytometry

Software Mentioned

ImageJ
Excel
BD FACS Diva
GraphPad
GraphPad Prism

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