Time-dependent cytotoxicity of dichloroacetate and metformin against Lewis lung carcinoma
DOI:
https://doi.org/10.32471/exp-oncology.2312-8852.vol-41-no-1.12432Keywords:
cytotoxicity, dichloroacetate, Lewis lung carcinoma, metformin, tumor cell migrationAbstract
Summary. The use of inhibitors of energy metabolism of malignant cells is a new promising trend in the treatment of cancer patients, based on one of the unique features of the malignant cell, namely the dominance of glycolysis over oxidative phosphorylation, even in the presence of oxygen, the so-called Warburg effect. Aim: To study time-dependent cytotoxicity of sodium dichloroacetate (DCA) and metformin (MTF) against metastatic tumor cells and action of these agents on tumor cell migration. Materials and Methods: In the study low metastatic LLC/R9 variant of Lewis lung carcinoma was used. The number of living cells in the cytotoxic test was evaluated using sulforhodamine B after 1, 2 and 3 days of cell incubation in vitro. The parameters of the sensitivity of tumor cells to the action of DCA and MTF in vitro were calculated using nonlinear and linear regression of experimental data. The effect of DCA and MTF on cellular motility in vitro was evaluated using a Boyden chamber by calculation of the number of cells that migrated to the bottom side of the filter within 3 days of incubation. The statistical analysis of the data was carried out with the use of descriptive methods, Student’s t-criterion, nonlinear, and linear regression analysis. Results: IC50 of DCA was found to be equal to 50.8 ± 7.6 mM at the first day of incubation with LLC/R9 cells and decreased by 1.9 (p < 0.05) and 2.1 (p < 0.05) times at the 2nd and 3rd days, respectively. Despite the almost identical ІC50 at the 2nd and 3rd days, an increase in the incubation period of cells with DCA for up to 3 days increased the C0 parameter, which reflects the maximum concentration of the agent that does not exhibit cytotoxic effects, by 93% (p < 0.05) compared to this at the 2nd day (16.2 ± 1.4 mM vs 8.4 ± 1.0 mM, correspondently). Unlike DCA, the LLC/R9 cell population was not homogeneous by the sensitivity to the action of MTF; at least at the 3rd day, an appearance of MTF-resistant subpopulation was observed, accounting for 35% of all cells. IC50 of MTF was equal to 12.1 ± 0.6 mM, and unlike DCA, this index progressively decreased at the 2nd and 3rd days by 1.4 (p < 0.05) and 9.3 times (p < 0.05) respectively. Action of DCA at a concentration of 25 mM alone and in combination with MTF at the concentrations of 0.1 mM and 0.7 mM resulted in an increase in cell migration by 65% (p < 0.05), 63% (p < 0.05) and 78.5% (p < 0.05), respectively. There was no significant effect of MTF on the tumor cell migration. Conclusions: The sensitivity of the metastatic Lewis lung carcinoma cells to the action of the modifiers of the energy metabolism increased significantly with an increase in the incubation period, apparently, primarily due to the shortage of nutrient substrates and, in particular, glucose, indicating the relevance of their combined use as well as with other agents, which promote the deficiency of glucose in the tumor microenvironment.
References
Michelakis ЕD, Webster L, Mackey JR. Dichloroacetate (DCA) as a potential metabolic-targeting therapy for cancer. Br J Cancer 2008; 99: 989–94.
Duan Y, Zhao X, Ren W, et al. Antitumor activity of dichloroacetate on C6 glioma cell: in vitro and in vivo evaluation. Onco Targets Ther 2013; 6: 189–98.
Ben Sahra I, Le Marchand-Brustel Y, Tanti JF, Bost F. Metformin in cancer therapy: a new perspective for an old antidiabetic drug? Mol Cancer Ther 2010; 9: 1092–9.
Ben Sahra I, Laurent K, Loubat A, et al. The antidiabetic drug metformin exerts an antitumoral effect in vitro and in vivo through a decrease of cyclin D1 level. Oncogene 2008; 27: 3576–86.
Kisfalvi K, Moro A, Sinnett-Smith J, et al. Metformin inhibits the growth of human pancreatic cancer xenografts. Pancreas 2013; 42: 781–5.
Drzewoski J, Drozdowska A, Sliwińska A. Do we have enough data to confirm the link between antidiabetic drug use and cancer development? Pol Arch Med Wewn 2011; 121: 81–7.
Zhou G, Myers R, Li Y et al. Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest 2001; 108: 1167–74.
Dowling RJ, Zakikhani M, Fantus IG, et al. Metformin inhibits mammalian target of rapamycin-dependent translation initiation in breast cancer cells. Cancer Res 2007; 67: 10804–12.
Zakikhani M, Dowling R, Fantus IG, et al. Metformin is an AMP kinase-dependent growth inhibitor for breast cancer cells. Cancer Res 2006; 66: 10269–73.
Ben Sahra I, Regazzetti C, Robert G, et al. Metformin, independent of AMPK, induces mTOR inhibition and cell-cycle arrest through REDD1. Cancer Res 2011; 71: 4366–72.
Chaudhary AK, Bhat TA, Kumar S, et al. Mitochondrial dysfunction-mediated apoptosis resistance associates with defective heat shock protein response in African-American men with prostate cancer. Br J Cancer 2016; 114: 1090–100.
Trinh SX, Nguyen HT, Saimuang K, et al. Metformin inhibits migration and invasion of cholangiocarcinoma cells. Asian Pac J Cancer Prev 2017; 18: 473–7.
Xintaropoulou C, Ward C, Wise A, et al. A comparative analysis of inhibitors of the glycolysis pathway in breast and ovarian cancer cell line models. Oncotarget 2015; 6: 25677–95.
Anemone A, Consolino L, Conti L, et al. In vivo evaluation of tumour acidosis for assessing the early metabolic response and onset of resistance to dichloroacetate by using magnetic resonance pH imaging. Intern J Cancer 2017; 51: 498–506.
Pyaskovskaya ON, Kolesnik DL, Fedorchuk AG, et al. Cytotoxic activity of metformin in vitro does not correlate with its antitumor action in vivo. Exp Oncol 2017; 39: 264–8.
Pyaskovskaya ON, Dasyukevich OI, Kolesnik DL, et al. Changes in VEGF level and tumor growth characteristics during Lewis lung carcinoma progression towards cis-DDP resistance. Exp Oncol 2007; 29: 197–202.
Vichai V, Kirtikara K. Sulforhodamine B colorimetric assay for cytoxicity screening. Nature Protocol 2006; 1: 1112–6.
Kolesnik DL, Pyaskovskaya ON, Tregubova NV, Solyanik GI. Lewis lung carcinoma variant with a high sensitivity to antitumor antiangiogenic therapy exhibits a high capacity for autophagy. Cytol Genet 2012; 46: 155–60.
Sanchez-Arago М, Chamorro M, Cuezva JM. Selection of cancer cells with repressed mitochondria triggers colon cancer progression. Carcinogenesis 2010; 31: 567–76.
LeBleu VS, O’Connell JT, Gonzalez Herrera KN, et al. PGC-1a mediates mitochondrial biogenesis and oxidative phosphorylation in cancer cells to promote metastasis. Nat Cell Biol 2014; 16: 992–1003.
Kolesnik DL, Pyaskovskaya ON, Boychuk IV, et al. Effect of dichloroacetate on Lewis lung carcinoma growth and metastasis. Exp Oncol 2015; 37: 126–9.
Schafer ZT, Grassian AR, Song L, et al. Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment. Nature 2009; 461: 109–13.
Piskounova E, Agathocleous M, Murphy MM, et al. Oxidative stress inhibits distant metastasis by human melanoma cells. Nature 2015; 527: 186–91.
Bonnet S, Archer SL, Allalunis-Turner J et al. A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell 2007; 11: 37–51.
Dai Y, Xiong X, Huang G et al. Dichloroacetate enhances adriamycin-induced hepatoma cell toxicity in vitro and in vivo by increasing reactive oxygen species levels. PLoS One 2014; 9: e92962.
Pyaskovskaya ON, Kolesnik DL, Kolobov AV, et al. Analysis of growth kinetics and proliferative heterogeneity of Lewis lung carcinoma cells growing as unfed culture. Exp Oncol 2008; 30: 269–75.
Stockwin LH, Yu SX, Borgel S, et al. Sodium dichloroacetate selectively targets cells with defects in the mitochondrial ETC. Int J Cancer 2010; 127: 2510–9.
De Preter G, Neveu MA, Danhier P, et al. Inhibition of the pentose phosphate pathway by dichloroacetate unravels a missing link between aerobic glycolysis and cancer cell proliferation. Oncotarget 2016; 7: 2910–20.
Ben Sahra I, Laurent K, Giuliano S, et al. Targeting cancer cell metabolism: the combination of metformin and 2-deoxyglucose induces p53-dependent apoptosis in prostate cancer cells. Cancer Res 2010; 70: 2465–75.
Bikas A, Jensen K, Patel A, et al. Glucose-deprivation increases thyroid cancer cells sensitivity to metformin. Endocr Relat Cancer 2015; 22: 919–32.
Zhuang Y, Chan DK, Haugrud AB, et al. Mechanisms by which low glucose enhances the cytotoxicity of metformin to cancer cells both in vitro and in vivo. PLoS One 2014; 9: e108444.
Menendez JA, Oliveras-Ferraros C, Cufí S, et al. Metformin is synthetically lethal with glucose withdrawal in cancer cells. Cell Cycle 2012; 11: 2782–92.
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