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PP242 suppresses cell proliferation, metastasis, and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway.

Xing X, Zhang L, Wen X, Wang X, Cheng X, Du H, Hu Y, Li L, Dong B, Li Z, Ji J - Anticancer Drugs (2014)

Bottom Line: Although preclinical work with rapalogs suggests potential in the treatment of gastric cancer, they have been less successful clinically.Furthermore, PP242 was found to decrease the tube formation and migration of human umbilical vein endothelial cells.These results show that PP242 suppresses cell proliferation and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway, which might be an effective novel therapeutic candidate against gastric cancer in the future.

View Article: PubMed Central - PubMed

Affiliation: Departments of aGastrointestinal Translational Research bGastrointestinal Surgery cTissue Bank dPathology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University, Beijing Cancer Hospital & Institute, Beijing, China.

ABSTRACT
Although preclinical work with rapalogs suggests potential in the treatment of gastric cancer, they have been less successful clinically. In this study, we report the impact of the investigational drug PP242, a potent and selective small-molecule active-site TORC1/2 kinase inhibitor, on tumor growth and metastasis. The antiproliferative effect of PP242 was assessed using the Cell Counting Kit-8 assay. The migration and invasion potential were analyzed using wound-healing and transwell assays, respectively. The Matrigel capillary tube formation assay was performed to mimic in-vivo angiogenesis. Immunoblotting and immunofluorescence were used to observe protein levels and distribution of actin fibers. Finally, p-mammalian target of rapamycin (mTOR) expression was detected on gastric cancer tissues using immunohistochemistry. First, PP242 potently inhibited cell proliferation in gastric cancer cell lines and in human endothelial cells in vitro at the IC50 ranged from 50 to 500 nmol/l. Then, an inhibitory effect of PP242 on metastasis was observed in gastric cancer cell AGS, along with the cytoskeletal rearrangements and suppression of the phosphorylation of PI3K downstream factors including AKT, mTOR, and P70S6K. Furthermore, PP242 was found to decrease the tube formation and migration of human umbilical vein endothelial cells. Using immunohistochemistry, we found that p-mTOR staining was observed in 41.8% (82/196) of gastric cancer tissues and correlated with depth of mural invasion, lymph node metastasis, tumor node metastasis stage, and vascular invasion. These results show that PP242 suppresses cell proliferation and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway, which might be an effective novel therapeutic candidate against gastric cancer in the future.

No MeSH data available.


Related in: MedlinePlus

Effect of PP242 on the motility capability of gastric cancer cell AGS and endothelial cell HUVEC. (a) Representative wound-healing images at 0 and 48 h. Wounds were created with a pipette tip in confluent monolayers. (b) Quantification of wound-healing rates. Data are mean±SD. *P<0.05, **P<0.01, ***P<0.001. HUVEC, human umbilical vein endothelial cell.
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Figure 2: Effect of PP242 on the motility capability of gastric cancer cell AGS and endothelial cell HUVEC. (a) Representative wound-healing images at 0 and 48 h. Wounds were created with a pipette tip in confluent monolayers. (b) Quantification of wound-healing rates. Data are mean±SD. *P<0.05, **P<0.01, ***P<0.001. HUVEC, human umbilical vein endothelial cell.

Mentions: We investigated the inhibitory effects of PP242 on the chemotactic motility of AGS and HUVEC cells using a wound-healing assay. Cells were carefully wounded and a continuous cell-free ‘wound’ region was observed. After 48 h incubation with increasing concentrations of PP242, the motility capacity of gastric cancer cells and human endothelial cells to move into the ‘wound’ region was significantly reduced in a dose-dependent manner (Fig. 2) as metastasis is one of most the critical events in tumor progression and migration is essential for endothelial cells to form blood vessels in angiogenesis and is necessary for tumor growth and metastasis. To examine the effect of PP242 on tumor invasion and metastasis (Fig. 3), we performed the invasion assay and the results were consistent with the observations obtained with the wound-healing assay. PP242 significantly inhibited AGS and HUVEC migration and invasion in a dose-dependent manner (Fig. 3, P<0.05).


PP242 suppresses cell proliferation, metastasis, and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway.

Xing X, Zhang L, Wen X, Wang X, Cheng X, Du H, Hu Y, Li L, Dong B, Li Z, Ji J - Anticancer Drugs (2014)

Effect of PP242 on the motility capability of gastric cancer cell AGS and endothelial cell HUVEC. (a) Representative wound-healing images at 0 and 48 h. Wounds were created with a pipette tip in confluent monolayers. (b) Quantification of wound-healing rates. Data are mean±SD. *P<0.05, **P<0.01, ***P<0.001. HUVEC, human umbilical vein endothelial cell.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222793&req=5

Figure 2: Effect of PP242 on the motility capability of gastric cancer cell AGS and endothelial cell HUVEC. (a) Representative wound-healing images at 0 and 48 h. Wounds were created with a pipette tip in confluent monolayers. (b) Quantification of wound-healing rates. Data are mean±SD. *P<0.05, **P<0.01, ***P<0.001. HUVEC, human umbilical vein endothelial cell.
Mentions: We investigated the inhibitory effects of PP242 on the chemotactic motility of AGS and HUVEC cells using a wound-healing assay. Cells were carefully wounded and a continuous cell-free ‘wound’ region was observed. After 48 h incubation with increasing concentrations of PP242, the motility capacity of gastric cancer cells and human endothelial cells to move into the ‘wound’ region was significantly reduced in a dose-dependent manner (Fig. 2) as metastasis is one of most the critical events in tumor progression and migration is essential for endothelial cells to form blood vessels in angiogenesis and is necessary for tumor growth and metastasis. To examine the effect of PP242 on tumor invasion and metastasis (Fig. 3), we performed the invasion assay and the results were consistent with the observations obtained with the wound-healing assay. PP242 significantly inhibited AGS and HUVEC migration and invasion in a dose-dependent manner (Fig. 3, P<0.05).

Bottom Line: Although preclinical work with rapalogs suggests potential in the treatment of gastric cancer, they have been less successful clinically.Furthermore, PP242 was found to decrease the tube formation and migration of human umbilical vein endothelial cells.These results show that PP242 suppresses cell proliferation and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway, which might be an effective novel therapeutic candidate against gastric cancer in the future.

View Article: PubMed Central - PubMed

Affiliation: Departments of aGastrointestinal Translational Research bGastrointestinal Surgery cTissue Bank dPathology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University, Beijing Cancer Hospital & Institute, Beijing, China.

ABSTRACT
Although preclinical work with rapalogs suggests potential in the treatment of gastric cancer, they have been less successful clinically. In this study, we report the impact of the investigational drug PP242, a potent and selective small-molecule active-site TORC1/2 kinase inhibitor, on tumor growth and metastasis. The antiproliferative effect of PP242 was assessed using the Cell Counting Kit-8 assay. The migration and invasion potential were analyzed using wound-healing and transwell assays, respectively. The Matrigel capillary tube formation assay was performed to mimic in-vivo angiogenesis. Immunoblotting and immunofluorescence were used to observe protein levels and distribution of actin fibers. Finally, p-mammalian target of rapamycin (mTOR) expression was detected on gastric cancer tissues using immunohistochemistry. First, PP242 potently inhibited cell proliferation in gastric cancer cell lines and in human endothelial cells in vitro at the IC50 ranged from 50 to 500 nmol/l. Then, an inhibitory effect of PP242 on metastasis was observed in gastric cancer cell AGS, along with the cytoskeletal rearrangements and suppression of the phosphorylation of PI3K downstream factors including AKT, mTOR, and P70S6K. Furthermore, PP242 was found to decrease the tube formation and migration of human umbilical vein endothelial cells. Using immunohistochemistry, we found that p-mTOR staining was observed in 41.8% (82/196) of gastric cancer tissues and correlated with depth of mural invasion, lymph node metastasis, tumor node metastasis stage, and vascular invasion. These results show that PP242 suppresses cell proliferation and angiogenesis of gastric cancer through inhibition of the PI3K/AKT/mTOR pathway, which might be an effective novel therapeutic candidate against gastric cancer in the future.

No MeSH data available.


Related in: MedlinePlus