Метаболические изменения в фибробластах при взаимодействии с опухолевыми клетками: НАД(Ф)Н, ФАД, цитоплазматический рН и пероксид водорода

I. N. Druzhkova; M. M. Lukina; V. V. Dudenkova; N. I. Ignatova; M. V. Shirmanova; E. V. Zagaynova

doi:10.25557/2310-0435.2019.04.33-42

I. N. Druzhkova Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0003-0293-3069
M. M. Lukina Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0003-1374-8571
V. V. Dudenkova Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0001-6761-5585
N. I. Ignatova Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0002-4570-9342
M. V. Shirmanova Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0002-3207-7227
E. V. Zagaynova Privolzhsky Research Medical University, Nizhny Novgorod, Russia http://orcid.org/0000-0003-2097-0525

DOI: https://doi.org/10.25557/2310-0435.2019.04.33-42

Keywords: cancer-associated fibroblasts, tumor microenvironment, energy metabolism, cytoplasmic pH, hydrogen peroxide

Abstract

Background. In the last decades, the role of microenvironment in carcinogenesis and therapeutic outcome has gained increasing attention. Cancer-associated fibroblasts (CAFs) have emerged as key players among stromal cells, which support tumor progression. In this process, a specific metabolic interplay forms between tumor cells and neighboring CAFs, and disconnection of this coupling is considered as a new therapeutic approach to anti-cancer treatment. The aim of this study was to investigate the dynamics of metabolic changes in fibroblasts during co-culturing with cancer cells. HAD(P)H and FAD, the endogenous metabolic cofactors, intracellular pH and concentration of hydrogen peroxide were used as indexes of the metabolic status.
Methods. The study was performed on human skin fibroblasts and human cervical cancer Hela cells. Fibroblasts were cultivated in a mono- or coculture with cancer cells for 5 days. The metabolic cofactors were evaluated by intensity and lifetime of their own fluorescence. Cytoplasmic pH and hydrogen peroxide level were measured using genetically encoded fluorescent sensors, SypHer2 and HyPer2, respectively.

Results. A more alkaline pH and increased concentration of hydrogen peroxide were observed in fibroblasts during cocultivation with cancer cells compared to the monoculture. In addition, the FAD / NAD(P)H redox-ratio was increased while relative contributions of NAD(P)H and FAD with open conformation were decreased in fibroblast during co-cultivation with cancer cells, which indicates development of the oxidative metabolic status.
Conclusion. The study confirmed the active metabolic interplay of fibroblasts with cancer cells. The dynamics and extent of the metabolic changes during this interplay suggested that affecting NAD(P)H- and FAD-depended reactions in the energy metabolism is a promising method for uncoupling this metabolic dialogue.