The adaptive regulation of thiamine pyrophosphokinase-1 facilitates malignant growth during supplemental thiamine conditions.

Hunter C Jonus,Bradley S Hanberry,Jaeah Kim,Long H Dang,Shivani Khatu,Michael G Bartlett,Hendrik Luesch,Jason A Zastre

doi:10.18632/oncotarget.26259

Hunter C Jonus, Bradley S Hanberry + Show 6 more

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https://doi.org/10.18632/oncotarget.26259

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Abstract

Supplemental levels of vitamin B1 (thiamine) have been implicated in tumor progression. Tumor cells adaptively up-regulate thiamine transport during hypoxic stress. Upon uptake, thiamine pyrophosphokinase-1 (TPK1) facilitates the rapid phosphorylation of thiamine into thiamine pyrophosphate (TPP). However, the regulation of TPK1 during hypoxic stress is undefined. Understanding how thiamine homeostasis changes during hypoxia will provide critical insight into the malignant advantage supplemental thiamine may provide cancer cells. Using Western blot analysis and RT-PCR, we have demonstrated the post-transcriptional up-regulation of TPK1 in cancer cells following hypoxic exposure. TPK1 expression was also adaptively up-regulated following alterations of redox status by chemotherapeutic and antioxidant treatments. Although TPK1 was functionally up-regulated by hypoxia, HPLC analysis revealed a reduction in intracellular TPP levels. This loss was reversed by treatment with cell-permeable antioxidants and corresponded with reduced ROS production and enhanced cellular proliferation during supplemental thiamine conditions. siRNA-mediated knockdown of TPK1 directly enhanced basal ROS levels and reduced tumor cell proliferation. These findings suggest that the adaptive regulation of TPK1 may be an essential component in the cellular response to oxidative stress, and that during supplemental thiamine conditions its expression may be exploited by tumor cells for a redox advantage contributing to tumor progression.

Highlights

Malignant cells experience elevated levels of reactive oxygen species (ROS) during tumor progression
These findings suggest that the adaptive regulation of thiamine pyrophosphokinase-1 (TPK1) may be an essential component in the cellular response to oxidative stress, and that during supplemental thiamine conditions its expression may be exploited by tumor cells for a redox advantage contributing to tumor progression
An increase in Ehrlich ascites tumor proliferation was observed in mice administered thiamine at 12.5 times the recommended daily allowance (RDA) [11]

Summary

Introduction

Malignant cells experience elevated levels of reactive oxygen species (ROS) during tumor progression. This multifactorial byproduct arises from events including enhanced metabolic activity, increased activity of ROS-producing enzymes, dysfunctional mitochondrial metabolism, activated immune responses, intratumoral hypoxia, and therapeutic intervention (i.e. chemotherapeutics, ionizing radiation) [1, 2]. Constitutive activation of the transcription factor Nuclear factor erythroid-2–related factor 2 (NFE2L2, NRF2) in tumor cells up-regulates levels of the endogenous glutathione machinery, promoting tumorigenicity [3]. The stabilization of the oncogenic transcription factor hypoxia-inducible factor-1α (HIF1A, HIF-1α) regulates the expression of pyruvate dehydrogenase kinase-1 (PDK1), which subsequently limits ROS production by phosphorylating pyruvate dehydrogenase (PDH) and restricting mitochondrial metabolism [4]. The stabilization of the oncogenic transcription factor hypoxia-inducible factor-1α (HIF1A, HIF-1α) regulates the expression of pyruvate dehydrogenase kinase-1 (PDK1), which subsequently limits ROS production by phosphorylating pyruvate dehydrogenase (PDH) and restricting mitochondrial metabolism [4]. siRNA-mediated silencing of HIF-1α increases intracellular ROS levels both in vitro and in vivo, highlighting HIF-1α’s critical role in ROS maintenance [5]

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