PFK15, a Small Molecule Inhibitor of PFKFB3, Induces Cell Cycle Arrest, Apoptosis and Inhibits Invasion in Gastric Cancer

Abstract

PFKFB3 (6-phosphofructo-2-kinase) synthesizes fructose 2,6-bisphosphate (F2,6P2), which is an allosteric activator of 6-phosphofructo-1-kinase (PFK-1), the rate-limiting enzyme of glycolysis. Overexpression of the PFKFB3 enzyme leads to high glycolytic metabolism, which is required for cancer cells to survive in the harsh tumor microenvironment. The objective of this study was to investigate the antitumor activity of PFK15 (1-(4-pyridinyl)-3-(2-quinolinyl)-2-propen-1-one), a small molecule inhibitor of PFKFB3, against gastric cancer and to explore its potential mechanisms. The effects of PFK15 on proliferation, apoptosis and cell cycle progression in gastric cancer cells were evaluated by cytotoxicity and apoptosis assays, flow cytometry, and western blotting. In addition, the invasion inhibition effects of PFK15 were measured by transwell invasion assay and western blot analysis, and a xenograft tumor model was used to verify the therapeutic effect of PFK15 in vivo. Results showed that PFK15 inhibited the proliferation, caused cell cycle arrest in G0/G1 phase by blocking the Cyclin-CDKs/Rb/E2F signaling pathway, and induced apoptosis through mitochondria in gastric cancer cells. Tumor volume and weight were also significantly reduced upon intraperitoneal injection with PFK15 at 25 mg/kg. In addition, PFK15 inhibited the invasion of gastric cancer cells by downregulating focal adhesion kinase (FAK) expression and upregulating E-cadherin expression. Taken together, our findings indicate that PFK15 is a promising anticancer drug for treating gastric cancer.

Long noncoding RNA AGPG regulates PFKFB3-mediated tumor glycolytic reprogramming

“…Recent studies revealed the unexpected role of PFKFB3 in promoting cell proliferation by regulating the expression of important cell cycle proteins: cyclin-dependent kinase-1 (CDK1) is upregulated, and p27 is downregulated, partially owing to the nuclear delivery of F-2,6-BP 10 . As reported, PFKFB3 inhibition is a promising modality for cancer treatment because it suppresses glycolysis, proliferation, and metastasis in cancer cells [11][12][13] .…”

Metabolic regulation of calcium pumps in pancreatic cancer: role of phosphofructokinase-fructose-bisphosphatase-3 (PFKFB3)

“…However, a possible explanation is that the steady-state ATP concentration within the cell is dependent not only on the ATP production rate but also on the ATP consumption rate. Given that PFK15 inhibits cell proliferation and is reported to arrest the cell cycle [26,30]; which are major ATP consuming processes, it is possible that reduced ATP production, due to inhibition of glycolysis, coincides with a corresponding reduced ATP consumption; such that steady-state ATP is maintained. Furthermore, in the context of inhibition of the PMCA, it is also possible that PFK15 inhibits a significant pool of PFKFB that resides in close proximity to the PMCA at the membrane, where it provides a privileged ATP supply to Indeed, in the present study, PFKFB3 was identified in the plasma membrane fraction, indicating localisation of a significant pool of PFKFB3 at the plasma membrane.…”

“…PFKFB3 is overexpressed in numerous cancers including PDAC [27,28]; this finding led to the development of the specific small-molecule inhibitor 3PO and then more potent 3PO derivatives: PFK15 and PFK158 [24]. PFK15 has been effective in reducing tumour cell growth in vitro and has also reduced tumour growth and metastasis in xenograft models [29][30][31]. Importantly, neither animal studies nor phase I trials has identified any major detrimental effects of PFK158 suggesting that there are no off-target effects and that inhibition of PFKFB3 is not ubiquitously harmful [32][33][34].…”

The molecular basis of targeting PFKFB3 as a therapeutic strategy against cancer

“…The role of PFK15 was also validated in HNSCC, the pharmacological inhibition of PFKFB3 result in suppression of tumor growth and alleviation of the lung metastasis in the xenograft mice models [ 36 ]. In addition to tumor cell proliferation suppression and cell apoptosis inducing, PFK15 was also found inhibiting the invasion of gastric cancer cells through downregulating focal adhesion kinase expression and upregulating E-cadherin expression [ 43 ]. To evaluate the efficacy and safety of the PFK15-based synthetic compound, PFK158, phase I clinical trials (NCT02044861) were implemented in 2014, and no serious adverse events were reported during observation for almost one year.…”