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

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Related in: MedlinePlus

Effect of PFK15 on cell viability, F2,6P2 production and glucose uptake.(a) Cells grown in exponential phase were exposed to the indicated concentrations of PFK15 and, after 24 h, the effects on viability were determined by trypan blue exclusion. *p<0.05, compared with the GES-1 groups. (b) Time-dependent effects of PFK15 on MKN45 and AGS cell viability under the concentration of 10 μM. *p<0.05, compared with the control group of MKN45 cell line; #p<0.05, compared with the control group of AGS cell line. (c,d) To identify acute effects on known metabolic effects of PFKFB3 inhibition, MKN45 and AGS cells were exposed to PFK15 for 12 h and the F2,6P2 and 2-Deoxyglucose uptake (Glucose Assay Kit) were measured. (e) The rescue effect of F2,6P2 on PFK15 induced cell proliferation suppression. The marked reduction of cell proliferation caused by 10 μmol/L of PFK15 in MKN45 or AGS cells could be partially rescued by F2,6P2 addition. Data are represented as the means ± SD from three independent experiments. *p<0.05, compared with controls.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5036843&req=5

pone.0163768.g001: Effect of PFK15 on cell viability, F2,6P2 production and glucose uptake.(a) Cells grown in exponential phase were exposed to the indicated concentrations of PFK15 and, after 24 h, the effects on viability were determined by trypan blue exclusion. *p<0.05, compared with the GES-1 groups. (b) Time-dependent effects of PFK15 on MKN45 and AGS cell viability under the concentration of 10 μM. *p<0.05, compared with the control group of MKN45 cell line; #p<0.05, compared with the control group of AGS cell line. (c,d) To identify acute effects on known metabolic effects of PFKFB3 inhibition, MKN45 and AGS cells were exposed to PFK15 for 12 h and the F2,6P2 and 2-Deoxyglucose uptake (Glucose Assay Kit) were measured. (e) The rescue effect of F2,6P2 on PFK15 induced cell proliferation suppression. The marked reduction of cell proliferation caused by 10 μmol/L of PFK15 in MKN45 or AGS cells could be partially rescued by F2,6P2 addition. Data are represented as the means ± SD from three independent experiments. *p<0.05, compared with controls.

Mentions: PFK15 has been reported to be a specific and potent small molecule antagonist of PFKFB3, and is able to suppress the proliferation of various cancer cells at relatively low concentrations [14]. We examined the effects of PFK15 on gastric cancer cells by trypan blue exclusion and observed a dose-dependent suppression of cell proliferation after 24 h on MKN45, AGS, and BGC823 gastric cancer cells (IC50: MKN45, 6.59±3.1 μmol/L; AGS, 8.54±2.7 μmol/L; BGC, 10.56±2.4 μmol/L) and a time-dependent inhibition over 48 h on MKN45 and AGS cells under a concentration of 10 μmol/L (Fig 1a and 1b). Moreover, PFK15 caused <10% inhibition on normal gastric cells (GES-1) viability indicating that PFK15 is a selective inhibitor with low cytotoxicity in normal cells.


PFK15, a Small Molecule Inhibitor of PFKFB3, Induces Cell Cycle Arrest, Apoptosis and Inhibits Invasion in Gastric Cancer
Effect of PFK15 on cell viability, F2,6P2 production and glucose uptake.(a) Cells grown in exponential phase were exposed to the indicated concentrations of PFK15 and, after 24 h, the effects on viability were determined by trypan blue exclusion. *p<0.05, compared with the GES-1 groups. (b) Time-dependent effects of PFK15 on MKN45 and AGS cell viability under the concentration of 10 μM. *p<0.05, compared with the control group of MKN45 cell line; #p<0.05, compared with the control group of AGS cell line. (c,d) To identify acute effects on known metabolic effects of PFKFB3 inhibition, MKN45 and AGS cells were exposed to PFK15 for 12 h and the F2,6P2 and 2-Deoxyglucose uptake (Glucose Assay Kit) were measured. (e) The rescue effect of F2,6P2 on PFK15 induced cell proliferation suppression. The marked reduction of cell proliferation caused by 10 μmol/L of PFK15 in MKN45 or AGS cells could be partially rescued by F2,6P2 addition. Data are represented as the means ± SD from three independent experiments. *p<0.05, compared with controls.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036843&req=5

pone.0163768.g001: Effect of PFK15 on cell viability, F2,6P2 production and glucose uptake.(a) Cells grown in exponential phase were exposed to the indicated concentrations of PFK15 and, after 24 h, the effects on viability were determined by trypan blue exclusion. *p<0.05, compared with the GES-1 groups. (b) Time-dependent effects of PFK15 on MKN45 and AGS cell viability under the concentration of 10 μM. *p<0.05, compared with the control group of MKN45 cell line; #p<0.05, compared with the control group of AGS cell line. (c,d) To identify acute effects on known metabolic effects of PFKFB3 inhibition, MKN45 and AGS cells were exposed to PFK15 for 12 h and the F2,6P2 and 2-Deoxyglucose uptake (Glucose Assay Kit) were measured. (e) The rescue effect of F2,6P2 on PFK15 induced cell proliferation suppression. The marked reduction of cell proliferation caused by 10 μmol/L of PFK15 in MKN45 or AGS cells could be partially rescued by F2,6P2 addition. Data are represented as the means ± SD from three independent experiments. *p<0.05, compared with controls.
Mentions: PFK15 has been reported to be a specific and potent small molecule antagonist of PFKFB3, and is able to suppress the proliferation of various cancer cells at relatively low concentrations [14]. We examined the effects of PFK15 on gastric cancer cells by trypan blue exclusion and observed a dose-dependent suppression of cell proliferation after 24 h on MKN45, AGS, and BGC823 gastric cancer cells (IC50: MKN45, 6.59±3.1 μmol/L; AGS, 8.54±2.7 μmol/L; BGC, 10.56±2.4 μmol/L) and a time-dependent inhibition over 48 h on MKN45 and AGS cells under a concentration of 10 μmol/L (Fig 1a and 1b). Moreover, PFK15 caused <10% inhibition on normal gastric cells (GES-1) viability indicating that PFK15 is a selective inhibitor with low cytotoxicity in normal cells.

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Related in: MedlinePlus