From the James Graham Brown Cancer Center, Division of Medical Oncology and Hematology, Department of Medicine, University of Louisville, Louisville, Kentucky 40202.
J Biol Chem. 2014 Mar 28;289(13):9440-8. doi: 10.1074/jbc.M113.529990. Epub 2014 Feb 10.
Estradiol (E2) administered to estrogen receptor-positive (ER(+)) breast cancer patients stimulates glucose uptake by tumors. Importantly, this E2-induced metabolic flare is predictive of the clinical effectiveness of anti-estrogens and, as a result, downstream metabolic regulators of E2 are expected to have utility as targets for the development of anti-breast cancer agents. The family of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4) control glycolytic flux via their product, fructose-2,6-bisphosphate (F26BP), which activates 6-phosphofructo-1-kinase (PFK-1). We postulated that E2 might promote PFKFB3 expression, resulting in increased F26BP and glucose uptake. We demonstrate that PFKFB3 expression is highest in stage III lymph node metastases relative to normal breast tissues and that exposure of human MCF-7 breast cancer cells to E2 causes a rapid increase in [(14)C]glucose uptake and glycolysis that is coincident with an induction of PFKFB3 mRNA (via ER binding to its promoter), protein expression and the intracellular concentration of its product, F26BP. Importantly, selective inhibition of PFKFB3 expression and activity using siRNA or a PFKFB3 inhibitor markedly reduces the E2-mediated increase in F26BP, [(14)C]glucose uptake, and glycolysis. Furthermore, co-treatment of MCF-7 cells with the PFKFB3 inhibitor and the anti-estrogen ICI 182,780 synergistically induces apoptotic cell death. These findings demonstrate for the first time that the estrogen receptor directly promotes PFKFB3 mRNA transcription which, in turn, is required for the glucose metabolism and survival of breast cancer cells. Importantly, these results provide essential preclinical information that may allow for the ultimate design of combinatorial trials of PFKFB3 antagonists with anti-estrogen therapies in ER(+) stage IV breast cancer patients.
雌二醇(E2)给予雌激素受体阳性(ER(+))乳腺癌患者会刺激肿瘤摄取葡萄糖。重要的是,这种 E2 诱导的代谢爆发可预测抗雌激素的临床疗效,因此,E2 的下游代谢调节剂有望成为开发抗乳腺癌药物的靶点。6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶(PFKFB1-4)家族通过其产物果糖-2,6-二磷酸(F26BP)控制糖酵解通量,F26BP 激活 6-磷酸果糖-1-激酶(PFK-1)。我们推测,E2 可能促进 PFKFB3 的表达,从而增加 F26BP 和葡萄糖摄取。我们证明,与正常乳腺组织相比,PFKFB3 的表达在 III 期淋巴结转移中最高,并且人 MCF-7 乳腺癌细胞暴露于 E2 会导致[(14)C]葡萄糖摄取和糖酵解的快速增加,这与 PFKFB3 mRNA 的诱导(通过 ER 与其启动子结合)、蛋白表达和其产物 F26BP 的细胞内浓度一致。重要的是,使用 siRNA 或 PFKFB3 抑制剂选择性抑制 PFKFB3 的表达和活性会显著降低 E2 介导的 F26BP、[(14)C]葡萄糖摄取和糖酵解的增加。此外,MCF-7 细胞与 PFKFB3 抑制剂和抗雌激素 ICI 182,780 联合处理会协同诱导细胞凋亡。这些发现首次证明雌激素受体直接促进 PFKFB3 mRNA 的转录,而这反过来又是乳腺癌细胞葡萄糖代谢和存活所必需的。重要的是,这些结果提供了重要的临床前信息,可能允许最终设计 PFKFB3 拮抗剂与 ER(+)期 IV 期乳腺癌患者的抗雌激素治疗的联合试验。