2-deoxy-D-Glucose Synergizes with Doxorubicin or L-Buthionine Sulfoximine to Reduce Adhesion and Migration of Breast Cancer Cells

Abstract

Background: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. Materials and Methods: T47D human breast cancer cells were treated with 2-deoxy-D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescentphalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. Results: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. Conclusions: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.

Combined Treatment with 2-Deoxy-D-Glucose and Doxorubicin Enhances the in Vitro Efficiency of Breast Cancer Radiotherapy

“…A study have showed that among breast cancer cell lines tested, SKBR3 was the most sensitive to the growth inhibitory effects of 2DG (Mustafa et al, 2015). In our study, the combination of 2DG and radiation, at different doses, caused cell killing and was significantly different from either treatment alone in both cell lines ( Figure 1B).…”

“…2DG-6-P is not a substrate for glucose-6-phosphate dehydrogenase or phosphohexoisomerase. Therefore, once formed, 2DG-6-P is not further metabolized, and, therefore, the output from glycolysis (ATP) and the pentose phosphate pathway gets reduced, and 2DG-6-P accumulates in the cell until dephosphorylated by phosphorylase (Maher et al, 2004;Pelicano et al, 2006;Dwarakanath, 2009;Aghaee et al, 2012;Mustafa et al, 2015). 2DG treatment resulted in intracellular ATP depletion as a consequence of glycolysis inhibition.…”

“…Meanwhile, studies on the metabolism pathways of cancer cells have demonstrated that accelerated glucose uptake for anaerobic glycolysis (Warburg effect) ( Mohanti et al, 1996;Pelicano et al, 2006;Gupta et al, 2009;Sinthupibulyakit et al, 2009;Mustafa et al, 2015), and the loss of regulation between glycolyticmetabolism and respiration (Simons et al, 2007), are the major metabolic changes found in malignant cells. In general, cancer cells have increased rates of glycolysis as well as pentose phosphate cycle activity, and slightly reduced rates of respiration (Simons et al, 2007;Aykin-Burns et al, 2009).…”

“…Glucose analogs have been found to profoundly inhibit glucose metabolism in cancer cells in vitro and in vivo (Gupta et al, 2009;Sinthupibulyakit et al, 2009;Zhang and Aft, 2009;Ahmad et al, 2010;Mustafa et al, 2015). Of the many glucose analogs which have been investigated, 2-Deoxy-D-Glucose (2DG) has been proven effective in the inhibition of tumor metabolism and ATP production (Gupta et al, 2009;Ahmad et al, 2010;Mustafa et al, 2015;Wang et al, 2015). Therefore, 2DG has suggested as an adjuvant of radiation therapy and chemotherapy both in vitro and in vivo (Gupta et al, 2009).…”

“…Analyzing of the radiomodifying effects of 2DG, observed in several tumor cell lines, revealed that the administration time of 2DG with respect to irradiation plays an important role in determining the effects (Dwarkanath et al, 2001;Dwarakanath, 2009;Gupta et al, 2009). It was shown that 2DG results in cessation of cell growth, in a dose dependent manner, and reduces adhesion and migration of breast cancer cells when combined with doxorubicin or L-Buthionine Sulfoximine (Mustafa et al, 2015). Also it was suggested that evaluation of glucose usage, lactate production, and energy status could be useful for predicting the responses of tumors to the combined treatment of radiation and 2DG (Dwarkanath et al, 2001).…”