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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.

No MeSH data available.


Mechanism of PFK15 induced apoptosis.(a) After treated with various concentrations of PFK15 for 24 h, MKN45 and AGS cells were collected and the protein changes were analyzed by western blot assay. (b) The optical density of Bcl-2 and Bax protein levels were quantified to controls by ImageJ software. PFK15 decreased the ratio of Bcl-2/Bax in a concentration-dependent manner. Columns, mean; bars, SD. *P<0.05, compared with controls.
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pone.0163768.g005: Mechanism of PFK15 induced apoptosis.(a) After treated with various concentrations of PFK15 for 24 h, MKN45 and AGS cells were collected and the protein changes were analyzed by western blot assay. (b) The optical density of Bcl-2 and Bax protein levels were quantified to controls by ImageJ software. PFK15 decreased the ratio of Bcl-2/Bax in a concentration-dependent manner. Columns, mean; bars, SD. *P<0.05, compared with controls.

Mentions: The involvement of apoptosis was further confirmed by changes in apoptosis associated proteins after PFK15 treatment. For example, caspase 3 and caspase 9 protein levels were downregulated at concentrations of 7 and 9 μmol/L of PFK15, whereas caspase 8 protein levels were unchanged in both MKN45 and AGS cells (Fig 5a). In addition, cleaved caspase-3 protein expressions were significantly increased by PFK15 at 9 μmol/L in both cell lines.


PFK15, a Small Molecule Inhibitor of PFKFB3, Induces Cell Cycle Arrest, Apoptosis and Inhibits Invasion in Gastric Cancer
Mechanism of PFK15 induced apoptosis.(a) After treated with various concentrations of PFK15 for 24 h, MKN45 and AGS cells were collected and the protein changes were analyzed by western blot assay. (b) The optical density of Bcl-2 and Bax protein levels were quantified to controls by ImageJ software. PFK15 decreased the ratio of Bcl-2/Bax in a concentration-dependent manner. Columns, mean; bars, SD. *P<0.05, compared with controls.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0163768.g005: Mechanism of PFK15 induced apoptosis.(a) After treated with various concentrations of PFK15 for 24 h, MKN45 and AGS cells were collected and the protein changes were analyzed by western blot assay. (b) The optical density of Bcl-2 and Bax protein levels were quantified to controls by ImageJ software. PFK15 decreased the ratio of Bcl-2/Bax in a concentration-dependent manner. Columns, mean; bars, SD. *P<0.05, compared with controls.
Mentions: The involvement of apoptosis was further confirmed by changes in apoptosis associated proteins after PFK15 treatment. For example, caspase 3 and caspase 9 protein levels were downregulated at concentrations of 7 and 9 μmol/L of PFK15, whereas caspase 8 protein levels were unchanged in both MKN45 and AGS cells (Fig 5a). In addition, cleaved caspase-3 protein expressions were significantly increased by PFK15 at 9 μmol/L in both cell lines.

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.