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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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Role of PFK15 in cell apoptosis.(a) MKN45 and AGS cells were treated with various concentrations of PFK15 and collected after 24 h. Then cells were analyzed by flow cytometry after stained with Annexin V and PI. The percentages of early apoptotic (Annexin V+/ PI-) and late apoptotic (Annexin V+/ PI+) cells were quantified. (b) PFK15 treatment significantly increased the percentage of apoptotic cells, 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.g004: Role of PFK15 in cell apoptosis.(a) MKN45 and AGS cells were treated with various concentrations of PFK15 and collected after 24 h. Then cells were analyzed by flow cytometry after stained with Annexin V and PI. The percentages of early apoptotic (Annexin V+/ PI-) and late apoptotic (Annexin V+/ PI+) cells were quantified. (b) PFK15 treatment significantly increased the percentage of apoptotic cells, 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 is important for the maintenance of an anti-apoptotic state [17]. Consistent with these results, treatment with PFK15 resulted in apoptosis in MKN45 and AGS cells, as indicated by flow cytometry using an Annexin V-FITC Apoptosis Detection kit (Fig 4a). A time related effect of PFK15 on cell apoptosis was also confirmed and a time-dependent increase in apoptotic cells (early and late apoptotic cells) was noted up to 48 h post-treatment (Fig 4b).


PFK15, a Small Molecule Inhibitor of PFKFB3, Induces Cell Cycle Arrest, Apoptosis and Inhibits Invasion in Gastric Cancer
Role of PFK15 in cell apoptosis.(a) MKN45 and AGS cells were treated with various concentrations of PFK15 and collected after 24 h. Then cells were analyzed by flow cytometry after stained with Annexin V and PI. The percentages of early apoptotic (Annexin V+/ PI-) and late apoptotic (Annexin V+/ PI+) cells were quantified. (b) PFK15 treatment significantly increased the percentage of apoptotic cells, 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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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5036843&req=5

pone.0163768.g004: Role of PFK15 in cell apoptosis.(a) MKN45 and AGS cells were treated with various concentrations of PFK15 and collected after 24 h. Then cells were analyzed by flow cytometry after stained with Annexin V and PI. The percentages of early apoptotic (Annexin V+/ PI-) and late apoptotic (Annexin V+/ PI+) cells were quantified. (b) PFK15 treatment significantly increased the percentage of apoptotic cells, 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 is important for the maintenance of an anti-apoptotic state [17]. Consistent with these results, treatment with PFK15 resulted in apoptosis in MKN45 and AGS cells, as indicated by flow cytometry using an Annexin V-FITC Apoptosis Detection kit (Fig 4a). A time related effect of PFK15 on cell apoptosis was also confirmed and a time-dependent increase in apoptotic cells (early and late apoptotic cells) was noted up to 48 h post-treatment (Fig 4b).

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