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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.

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Related in: MedlinePlus

Role of PFK15 in cell cycle progression.(a) MKN45 and AGS cells were fixed and stained by PI following dedicated concentrations of PFK15 treatment after 24 h. Then the cell cycle distribution was analyzed using flow cytometer. PFK15 treatment remarkably prevented MKN45 cells from proceeding to S phase, thus resulting in cell cycle arrest at G0/G1 phase. (b) PFK15 treatment significantly increased the percentage of cells in G1 phase, indicating a time dependent effect on tested cells up to 48 h post-treatment. Data were analyzed by means ± SD and were representative of three independent experiments. *P<0.05, compared with controls.
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pone.0163768.g002: Role of PFK15 in cell cycle progression.(a) MKN45 and AGS cells were fixed and stained by PI following dedicated concentrations of PFK15 treatment after 24 h. Then the cell cycle distribution was analyzed using flow cytometer. PFK15 treatment remarkably prevented MKN45 cells from proceeding to S phase, thus resulting in cell cycle arrest at G0/G1 phase. (b) PFK15 treatment significantly increased the percentage of cells in G1 phase, indicating a time dependent effect on tested cells up to 48 h post-treatment. Data were analyzed by means ± SD and were representative of three independent experiments. *P<0.05, compared with controls.

Mentions: PFKFB3 expression has been shown to be important for cell cycle progression [17, 19]. For example, PFKFB3-knockdown cells resulted in an increased percentage of cells in the S phase of the cell cycle [20]. Therefore, we investigated the role of PFK15 in the cell cycle changes in MKN45 and AGS cells using flow cytometry analysis, as described in methods. Treatment with PFK15 for 24 h caused cell cycle arrest at the G0/G1 phase in both cell lines (Fig 2a). The percent of cells in the G0/G1 phase significantly increased from 50.7% in control group to 79.5% (P<0.05) after 9 μmol/L of PFK15 treatment in MKN45 cells and 54.1% to 72.5% in AGS cells, respectively. Simultaneously, the proportion of cells in S phase decreased. A time related effect of PFK15 on cell cycle progression was also confirmed and a time-dependent increase in G1 phase cells was noted up to 48 h post-treatment (Fig 2b).


PFK15, a Small Molecule Inhibitor of PFKFB3, Induces Cell Cycle Arrest, Apoptosis and Inhibits Invasion in Gastric Cancer
Role of PFK15 in cell cycle progression.(a) MKN45 and AGS cells were fixed and stained by PI following dedicated concentrations of PFK15 treatment after 24 h. Then the cell cycle distribution was analyzed using flow cytometer. PFK15 treatment remarkably prevented MKN45 cells from proceeding to S phase, thus resulting in cell cycle arrest at G0/G1 phase. (b) PFK15 treatment significantly increased the percentage of cells in G1 phase, indicating a time dependent effect on tested cells up to 48 h post-treatment. Data were analyzed by means ± SD and were representative of three independent experiments. *P<0.05, compared with controls.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5036843&req=5

pone.0163768.g002: Role of PFK15 in cell cycle progression.(a) MKN45 and AGS cells were fixed and stained by PI following dedicated concentrations of PFK15 treatment after 24 h. Then the cell cycle distribution was analyzed using flow cytometer. PFK15 treatment remarkably prevented MKN45 cells from proceeding to S phase, thus resulting in cell cycle arrest at G0/G1 phase. (b) PFK15 treatment significantly increased the percentage of cells in G1 phase, indicating a time dependent effect on tested cells up to 48 h post-treatment. Data were analyzed by means ± SD and were representative of three independent experiments. *P<0.05, compared with controls.
Mentions: PFKFB3 expression has been shown to be important for cell cycle progression [17, 19]. For example, PFKFB3-knockdown cells resulted in an increased percentage of cells in the S phase of the cell cycle [20]. Therefore, we investigated the role of PFK15 in the cell cycle changes in MKN45 and AGS cells using flow cytometry analysis, as described in methods. Treatment with PFK15 for 24 h caused cell cycle arrest at the G0/G1 phase in both cell lines (Fig 2a). The percent of cells in the G0/G1 phase significantly increased from 50.7% in control group to 79.5% (P<0.05) after 9 μmol/L of PFK15 treatment in MKN45 cells and 54.1% to 72.5% in AGS cells, respectively. Simultaneously, the proportion of cells in S phase decreased. A time related effect of PFK15 on cell cycle progression was also confirmed and a time-dependent increase in G1 phase cells was noted up to 48 h post-treatment (Fig 2b).

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