REPROGRAMMING OF GLUCOSE METABOLISM IN HUMAN BREAST CANCER CELLS AFTER CO-CULTIVATION WITH BIFIDOBACTERIUM ANIMALIS
DOI:
https://doi.org/10.15407/exp-oncology.2025.01.003Keywords:
breast cancer, Bifidobacterium, glucose, lactate, STAT6, GLUT1Abstract
Background. The ability to reorganize metabolic processes is one of the key properties of malignant cells necessary to ensure high energy needs, survival, proliferation, metastasis, and resistance to anticancer drugs. Lactic acid bacteria, in particular Bifidobacteria, are important elements of the tumor microenvironment in breast cancer (BC) and, as active lactate producers, can influence the metabolic phenotype of malignant cells. Aim. To study the effect of B. animalis on some components of glucose metabolism pathways and the expression of proteins associated with this process in human BC cells of different molecular subtypes. Materials and Methods. The study was performed on human BC cells of the T-47D, MCF-7 (luminal subtype), and MDA-MB-231 (basal subtype) lines and live culture of Bifidobacterium animalis subsp. lactis (B. animalis). A colorimetric enzymatic technique, flow cytometry, immunocytochemical analysis, and cell viability trypan blue exclusion assay were used in the study. Results. Co-cultivation of BC cells with B. animalis resulted in a significant (p < 0.05) increase in the glucose consumption rate by 1.2—4.7 times, lactate production by 15—115%, and LDH activity by 15—160% in BC cells compared to control cells. The most pronounced changes were observed in BC cells of the luminal subtype where they were accompanied by an increase in the expression of the GLUT1 glucose transporter by 30—80% compared to control cells. Also, after co-cultivation with B. animalis, we detected an increased expression of the STAT6 transcription factor in BC cells of all three lines. Conclusions. Co-cultivation of BC cells with B. animalis is accompanied by an increase in glycolysis. B. animalis affected not only the biochemical components of the glucose metabolism pathway but also the expression levels of STAT6, GLUT1, and insulin receptor.
References
Siegel RL, Giaquinto AN, Jemal A. Cancer statistics, 2024. CA Cancer J Clin. 2024;74:12-49. https://doi.org/10.3322/ caac.21820
Chekhun V, Martynyuk O, Lukianova Ye, et al. Features of breast cancer in patients of young age: search for di- agnosis optimization and personalized treatment. Exp Oncol. 2023;45(2):139-150. https://doi.org/10.15407/exp- oncology.2023.02.139
Lykhova O, Zavelevich M, Philchenkov A, et al. Does insulin make breast cancer cells resistant to doxorubicin tox- icity? Naunyn Schmiedebergs Arch Pharmacol. 2023;396:3111-3122. https://doi.org/10.1007/s00210-023-02516-3
Proietto M, Crippa M, Damiani C, et al. Tumor heterogeneity: preclinical models, emerging technologies, and fu- ture applications. Front Oncol. 2023;13:1164535. https://doi.org/10.3389/fonc.2023.1164535
Kim J, DeBerardinis RJ. Mechanisms and implications of metabolic heterogeneity in cancer. Cell Metab. 2019;30:434- 446. https://doi.org/10.1016/j.cmet.2019.08.013
Gandhi N, Das G. Metabolic reprogramming in breast cancer and its therapeutic implications. Cells. 2019;8:89. https://doi.org/10.3390/cells8020089
Liu S, Zhang X, Wang W, et al. Metabolic reprogramming and therapeutic resistance in primary and metastatic breast cancer. Mol Cancer. 2024;23:261. https://doi.org/10.1186/s12943-024-02165-x
Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009;324:1029-1033. https://doi.org/10.1126/science.1160809
Lin X, Xiao Z, Chen T, et al. Glucose metabolism on tumor plasticity, diagnosis, and treatment. Front Oncol. 2020;10:317. https://doi.org/10.3389/fonc.2020.00317
Mikó E, Kovács T, Sebő É, et al. Microbiome—microbial metabolome—cancer cell interactions in breast cancer— familiar, but unexplored. Cells. 2019;8:293. https://doi.org/10.3390/cells8040293
Tzeng A, Sangwan N, Jia M, et al. Human breast microbiome correlates with prognostic features and immunologi- cal signatures in breast cancer. Genome Med. 2021;13:60. https://doi.org/10.1186/s13073-021-00874-2
Nowak A, Paliwoda A, Błasiak J. Anti-proliferative, pro-apoptotic and anti-oxidative activity of lactobacillus and bi- fidobacterium strains: a review of mechanisms and therapeutic perspectives. Crit Rev Food Sci Nutr. 2019;59:3456- 3467. https://doi.org/10.1080/10408398.2018.1494539
Sharma D, Gajjar D, Seshadri S. Understanding the role of gut microfloral bifidobacterium in cancer and its poten- tial therapeutic applications. Microbiome Res Rep. 2023;3:3. https://doi.org/10.20517/mrr.2023.51
Mizuta M, Endo I, Yamamoto S, et al. Perioperative supplementation with bifidobacteria improves postopera- tive nutritional recovery, inflammatory response, and fecal microbiota in patients undergoing colorectal surgery: a prospective, randomized clinical trial. Biosci Microbiota Food Health. 2016;35:77-87. https://doi.org/10.12938/ bmfh.2015-017
Zaharuddin L, Mokhtar NM, Muhammad Nawawi KN, Raja Ali RA. A randomized double-blind placebo- controlled trial of probiotics in post-surgical colorectal cancer. BMC Gastroenterol. 2019;19:131. https://doi. org/10.1186/s12876-019-1047-4
Procaccianti G, Roggiani S, Conti G, et al. Bifidobacterium in anticancer immunochemotherapy: friend or foe?
Microbiome Res Rep. 2023;2:2. https://doi.org/10.20517/mrr.2023.23
Xing W, Li X, Zhou Y, et al. Lactate metabolic pathway regulates tumor cell metastasis and its use as a new thera- peutic target. Explor Med. 2023:541-559. https://doi.org/10.37349/emed.2023.00160
CapesDavis A, Freshney RI. Freshney’s Culture of Animal Cells: A Manual of Basic Technique and Specialized Ap- plications. Ed. by Robert J. Geraghty and Raymond W. Nims. Eighth edition. Hoboken, NJ: Wiley Blackwell, 2021.
Riisom M, Jamieson SMF, Hartinger CG. Critical evaluation of cell lysis methods for metallodrug studies in cancer cells. Metallomics. 2023;15:mfad048. https://doi.org/10.1093/mtomcs/mfad048
Solyanik GI, Kolesnik DL, Prokhorova IV, et al. Mitochondrial dysfunction significantly contributes to the sensi- tivity of tumor cells to anoikis and their metastatic potential. Heliyon. 2024;10:e32626. https://doi.org/10.1016/j. heliyon.2024.e32626
Detre S, Saclani Jotti G, Dowsett M. A quickscore method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas. J Clin Pathol. 1995;48:876-878. https://doi.org/10.1136/jcp.48.9.876
Martin SD, McGee SL. A systematic flux analysis approach to identify metabolic vulnerabilities in human breast cancer cell lines. Cancer Metab. 2019;7:12. https://doi.org/10.1186/s40170-019-0207-x
Vella V, Milluzzo A, Scalisi NM, et al. Insulin receptor isoforms in cancer. Int J Mol Sci. 2018;19:3615. https://doi. org/10.3390/ijms19113615
PérezTomás R, PérezGuillén I. Lactate in the tumor microenvironment: an essential molecule in cancer progression and treatment. Cancers. 2020;12:3244. https://doi.org/10.3390/cancers12113244
Barron CC, Bilan PJ, Tsakiridis T, Tsiani E. Facilitative glucose transporters: implications for cancer detection, prognosis and treatment. Metabolism. 2016;65:124-139. https://doi.org/10.1016/j.metabol.2015.10.007
Schroder AJ, Pavlidis P, Arimura A, Capece D, Rothman PB. Cutting edge: STAT6 serves as a positive and neg- ative regulator of gene expression in IL4-stimulated B lymphocytes. J Immunol. 2002;168:996-1000. https://doi. org/10.4049/jimmunol.168.3.996
Hönigova K, Navratil J, Peltanova B, et al. Metabolic tricks of cancer cells. Biochim Biophys Acta Rev Cancer. 2022;1877:188705. https://doi.org/10.1016/j.bbcan.2022.188705
D’Amico F, Perrone AM, Rampelli S, et al. Gut microbiota dynamics during chemotherapy in epithelial ovarian can- cer patients are related to therapeutic outcome. Cancers. 2021;13:3999. https://doi.org/10.3390/cancers13163999
Arnone AA, Tsai YT, Cline JM, et al. Endocrine-targeting therapies shift the breast microbiome to reduce estrogen receptor α breast cancer risk. Cell Rep Med. 2025;6:101880. https://doi.org/10.1016/j.xcrm.2024.101880
Niamah AK, AlSahlany STG, Verma DK, et al. Emerging lactic acid bacteria bacteriocins as anticancer and antitu- mor agents for human health. Heliyon. 2024;10:e37054. https://doi.org/10.1016/j.heliyon.2024.e37054
Asadollahi P, Ghanavati R, Rohani M, et al. Anticancer effects of bifidobacterium species in colon cancer cells and a mouse model of carcinogenesis. PLoS One. 2020;15:e0232930. https://doi.org/10.1371/journal.pone.0232930
Kozak T, Lykhova O. Suppression of proliferation and increased of proapoptotic proteins expression in human breast cancer cells after their cocultivation with bifidobacterium animalis in vitro. Oncology. 2024;25:29-37 (in Ukrainian). https://doi.org/10.15407/oncology.2024.01.029
Wang Z, Jiang Q, Dong C. Metabolic reprogramming in triple-negative breast cancer. Cancer Biol Med. 2020;17:44- 59. https://doi.org/10.20892/j.issn.2095-3941.2019.0210
Cordani M, Michetti F, Zarrabi A, et al. The role of glycolysis in tumorigenesis: from biological aspects to therapeu- tic opportunities. Neoplasia. 2024;58:101076. https://doi.org/10.1016/j.neo.2024.101076.
Pavlova NN, Thompson CB. The emerging hallmarks of cancer metabolism. Cell Metab. 2016;23:27-45. https://doi. org/10.1016/j.cmet.2015.12.006
Wu C, Khan SA, Lange AJ. Regulation of glycolysis—role of insulin. Exp Gerontol. 2005;40:894-899. https://doi. org/10.1016/j.exger.2005.08.002
Krishnapuram R, KirkBallard H, Dhurandhar EJ, et al. Insulin receptor-independent upregulation of cellular glu- cose uptake. Int J Obes. 2013;37:146-153. https://doi.org/10.1038/ijo.2012.6
Wu Q, BaAlawi W, Deblois G, et al. GLUT1 inhibition blocks growth of RB1-positive triple negative breast cancer.
Nat Commun. 2020;11:4205. https://doi.org/10.1038/s41467-020-18020-8
Medina RA, Southworth R, Fuller W, Garlick PB. Lactate-induced translocation of GLUT1 and GLUT4 is not medi- ated by the phosphatidylinositol-3-kinase pathway in the rat heart. Basic Res Cardiol. 2002;97:168-176. https://doi. org/10.1007/s003950200008
Ghanavat M, Shahrouzian M, Deris Zayeri Z, et al. Digging deeper through glucose metabolism and its regulators in cancer and metastasis. Life Sci. 2021;264:118603. https://doi.org/10.1016/j.lfs.2020.118603
Zhang Y, Li Q, Huang Z, et al. Targeting glucose metabolism enzymes in cancer treatment: current and emerging strategies. Cancers. 2022;14:4568. https://doi.org/10.3390/cancers14194568
Salazar G. NADPH oxidases and mitochondria in vascular senescence. Int J Mol Sci. 2018;19:1327. https://doi. org/10.3390/ijms19051327
Mele L, La Noce M, Paino F, et al. Glucose-6-phosphate dehydrogenase blockade potentiates tyrosine kinase inhibi- tor effect on breast cancer cells through autophagy perturbation. J Exp Clin Cancer Res. 2019;38:160. https://doi. org/10.1186/s13046-019-1164-5
Dufort FJ, Bleiman BF, Gumina MR, et al. Cutting edge: IL4-mediated protection of primary B lymphocytes from apoptosis via Stat6-dependent regulation of glycolytic metabolism. J Immunol. 2007;179:4953-4957. https://doi. org/10.4049/jimmunol.179.8.4953
Venmar KT, Kimmel DW, Cliffel DE, Fingleton B. IL4 receptor α mediates enhanced glucose and glutamine me- tabolism to support breast cancer growth. Biochim Biophys Acta Mol Cell Res. 2015;1853:1219-1228. https://doi. org/10.1016/j.bbamcr.2015.02.020
Wei M, Liu B, Gu Q, et al. Stat6 cooperates with Sp1 in controlling breast cancer cell proliferation by modulating the expression of p21Cip1/WAF1 and p27Kip1. Cell Oncol. 2013;36:79-93. https://doi.org/10.1007/s13402-012-0115-3
Verhoeven Y, Tilborghs S, Jacobs J, et al. The potential and controversy of targeting STAT family members in cancer.
Semin Cancer Biol. 2020;60:41-56. https://doi.org/10.1016/j.semcancer.2019.10.002
Wagner W, Ciszewski W, Kania K, Dastych J. Lactate stimulates IL4 and IL13 production in activated HuT78 T lym- phocytes through a process that involves monocarboxylate transporters and protein hyperacetylation. J Interferon Cytokine Res. 2016;36:317-327. https://doi.org/10.1089/jir.2015.0086
Lunt SY, Vander Heiden MG. Aerobic glycolysis: meeting the metabolic requirements of cell proliferation. Annu Rev Cell Dev Biol. 2011;27:441-464. https://doi.org/10.1146/annurev-cellbio-092910-154237
Yasuhara N, Eguchi Y, Tachibana T, et al. Essential role of active nuclear transport in apoptosis. Genes Cells. 1997;2:55-64. https://doi.org/10.1046/j.1365-2443.1997.1010302.x
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