The impact of berberine on the energy metabolism of cancer cells and CAFs in non-small lung cancer.
Authors: Mezhevova Irina V. , Kit Oleg I. , Filippova Svetlana Yu , Sitkovskaya Anastasia O. , Gnennaya Nadezhda V. , Shamova Tatiana V. , Timofeeva Sofia V. , Novikova Inna A. , Zlatnik Elena Yu , Enin Yaroslav S. , Burov Oleg N. , Kharagezov Dmitry A. , Stateshny Oleg N. , Milakin Anton G. , Leyman Igor A. , Mirzoyan Ellada A. , Gappoeva Madina A. , Lazutin Yuriy N. , Vitkovskaya Victoria N. , Iozefi Kristian D. 
Published: June 2022

e15097 Background: The energy metabolism of tumor cells and infiltrating stromal cells is a promising target in anticancer therapy. One of the mechanisms of antitumor activity of berberine is its ability to suppress oxidative phosphorylation in cancer cells. However, there is relatively little data on how berberine affects stromal cells compared to cancer cells. This study assessed the effect of berberine on the energy metabolism in non-small cell lung cancer (NSCLC) cell, as well as cancer-associated fibroblasts (CAFs) of the same localization. Methods: Cells of CAFs, NSCLC primary cell culture and permanent lung cancer culture H1299 were seeded in an amount of 2*104 per well in a Seahorse XFp Analyzer plate (Agilent, USA) in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA). After cell adhesion berberine at a concentration of 2.5 µM or an equal amount of medium without berberine was added. For each cell culture experimental and control wells were set up in 3 repetitions. After 24 hours of cultivation, the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured using the Seahorse XFp Analyzer (Agilent, USA) using the Seahorse XFp Cell Energy Phenotype Test (Agilent, USA), with 2 µM FCCP and 1 µM oligomycin. Results: Cultivation with berberine resulted in a significant (α = 0.05, df = 4) decrease in baseline OCR in all cultures, which was more pronounced in CAFs. The decrease in the baseline OCR in CAFs was 20.65±5%, in the primary culture 17.56±2.1% and in the permanent culture of H1299 8.29±1.1% compared to the control samples. At the same time, the maximum level of OCR also significantly decreased in CAFs compared to the control by 47.13±6.2% and in the H1299 culture by 14.9±3.1% (α = 0.05, df = 4). However, in the primary culture of NSCLC, the decrease in maximum OCR compared to the control was only 3.87±1.2% and was not significant. Both primary NSCLC culture and CAFs exhibited an increase in baseline glycolysis in response to berberine addition. In the primary cell culture ECAR increased by 19.7±2.3%, and in the culture of fibroblasts by 18.9±3.8% compared to the control. On the contrary, the baseline level of glycolysis in the permanent cell line H1299 decreased, which can be judged by a decrease in ECAR by 32.8±5.9% compared with the control. ECAR level changes were significant in all cultures at the accepted level of significance (α = 0.05, df = 4). Conclusions: Berberine causes oxidative phosphorylation inhibition in both cancerous cells and CAFs. Nevertheless, a compensatory increase in the level of glycolysis is only observed in primary cell cultures of NSCLC and CAFs.