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Saracatinib impairs the peritoneal dissemination of diffuse-type gastric carcinoma cells resistant to Met and fibroblast growth factor receptor inhibitors.

Yamaguchi H, Takanashi M, Yoshida N, Ito Y, Kamata R, Fukami K, Yanagihara K, Sakai R - Cancer Sci. (2014)

Bottom Line: Saracatinib also effectively impaired peritoneal dissemination of Met-independent and FGFR2-independent SGC cells.Moreover, DGC cell lines exhibited nearly mutually exclusive susceptibility to Met, FGFR and Src inhibitors.These results suggest that DGC have distinct sensitivities to molecular target drugs and that targeting Src is beneficial in the treatment of DGC insensitive to Met and FGFR inhibition.

View Article: PubMed Central - PubMed

Affiliation: Division of Metastasis and Invasion Signaling, National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan.

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44As3 and 58As9 scirrhous gastric carcinoma (SGC) cells have different sensitivities to Met inhibitors. (a, b) 44As3 and 58As9 cells were treated with 100 or 300 nM of Met inhibitors PHA-665752 (PHA) or JNJ-3887760 (JNJ) for 2 h and subjected to immunoblot analyses. (c) Phase contrast micrographs of cells treated with Met inhibitors for 1 day. (d) Cells were cultured in the presence of 10 or 0.1% serum for 1 day and then stimulated with 100 ng/mL hepatocyte growth factor (HGF) for 10 min. The cells were then analyzed with immunoblotting. (e) Viability of cells cultured in the presence of various concentrations of serum for 3 days. Bars, SD (n = 4). *P < 0.005 and **P < 0.0005 by Student's t-test. (f) 44As3 and 58As9 cells were inoculated intraperitoneally into nude mice, and DMSO (vehicle) or the Met inhibitor PHA was administered. Representative macroscopic views of mesentery tumor nodules are shown. (g) The number of mesentery tumor nodules (>1 mm in diameter). Bars, SEM (n = 11 for DMSO and 13 for PHA in 44As3; n = 10 for DMSO and PHA in 58As9). *P < 0.001 by the Mann–Whitney test.
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fig02: 44As3 and 58As9 scirrhous gastric carcinoma (SGC) cells have different sensitivities to Met inhibitors. (a, b) 44As3 and 58As9 cells were treated with 100 or 300 nM of Met inhibitors PHA-665752 (PHA) or JNJ-3887760 (JNJ) for 2 h and subjected to immunoblot analyses. (c) Phase contrast micrographs of cells treated with Met inhibitors for 1 day. (d) Cells were cultured in the presence of 10 or 0.1% serum for 1 day and then stimulated with 100 ng/mL hepatocyte growth factor (HGF) for 10 min. The cells were then analyzed with immunoblotting. (e) Viability of cells cultured in the presence of various concentrations of serum for 3 days. Bars, SD (n = 4). *P < 0.005 and **P < 0.0005 by Student's t-test. (f) 44As3 and 58As9 cells were inoculated intraperitoneally into nude mice, and DMSO (vehicle) or the Met inhibitor PHA was administered. Representative macroscopic views of mesentery tumor nodules are shown. (g) The number of mesentery tumor nodules (>1 mm in diameter). Bars, SEM (n = 11 for DMSO and 13 for PHA in 44As3; n = 10 for DMSO and PHA in 58As9). *P < 0.001 by the Mann–Whitney test.

Mentions: The IC50 values of Met inhibitors in 44As3 and 58As9 cells were approximately 1500 and 19 nM for PHA-665752, >10 000 and 6.9 nM for JNJ-38877605, and 2600 and 21 nM for PF-2341066, respectively (Fig. S2a). Met inhibitors significantly blocked phosphorylation of Met in both 44As3 and 58As9 cells (Fig.2a). Nonetheless, overall protein tyrosine phosphorylation and phosphorylation of ERK, Akt and Stat3 were significantly decreased only in 58As9 cells (Fig.2a,b). Interestingly, Src phosphorylation was rather increased in 58As9 cells upon Met inhibition, whereas it was unchanged in 44As3 cells (Fig.2a). Similar results were obtained with PF-2341066 (Fig. S2b). Met inhibitor treatment also induced marked changes in cell morphology in 58As9 but not in 44As3 cells: rounded morphology was converted to flattened and adherent phenotypes (Fig.2c). Serum starvation reduced and following HGF stimulation increased Met phosphorylation in 44As3 cells, whereas Met was constitutively phosphorylated in 58As9 cells (Fig.2d). Consequently, growth of 44As3 but not of 58As9 cells was sensitive to serum starvation (Fig.2e). These results indicate that overexpressed Met proteins confer the capability for serum-independent growth to 58As9 cells via activation of downstream signaling pathways, and, therefore, that 58As9 cells are susceptible to Met inhibitors.


Saracatinib impairs the peritoneal dissemination of diffuse-type gastric carcinoma cells resistant to Met and fibroblast growth factor receptor inhibitors.

Yamaguchi H, Takanashi M, Yoshida N, Ito Y, Kamata R, Fukami K, Yanagihara K, Sakai R - Cancer Sci. (2014)

44As3 and 58As9 scirrhous gastric carcinoma (SGC) cells have different sensitivities to Met inhibitors. (a, b) 44As3 and 58As9 cells were treated with 100 or 300 nM of Met inhibitors PHA-665752 (PHA) or JNJ-3887760 (JNJ) for 2 h and subjected to immunoblot analyses. (c) Phase contrast micrographs of cells treated with Met inhibitors for 1 day. (d) Cells were cultured in the presence of 10 or 0.1% serum for 1 day and then stimulated with 100 ng/mL hepatocyte growth factor (HGF) for 10 min. The cells were then analyzed with immunoblotting. (e) Viability of cells cultured in the presence of various concentrations of serum for 3 days. Bars, SD (n = 4). *P < 0.005 and **P < 0.0005 by Student's t-test. (f) 44As3 and 58As9 cells were inoculated intraperitoneally into nude mice, and DMSO (vehicle) or the Met inhibitor PHA was administered. Representative macroscopic views of mesentery tumor nodules are shown. (g) The number of mesentery tumor nodules (>1 mm in diameter). Bars, SEM (n = 11 for DMSO and 13 for PHA in 44As3; n = 10 for DMSO and PHA in 58As9). *P < 0.001 by the Mann–Whitney test.
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fig02: 44As3 and 58As9 scirrhous gastric carcinoma (SGC) cells have different sensitivities to Met inhibitors. (a, b) 44As3 and 58As9 cells were treated with 100 or 300 nM of Met inhibitors PHA-665752 (PHA) or JNJ-3887760 (JNJ) for 2 h and subjected to immunoblot analyses. (c) Phase contrast micrographs of cells treated with Met inhibitors for 1 day. (d) Cells were cultured in the presence of 10 or 0.1% serum for 1 day and then stimulated with 100 ng/mL hepatocyte growth factor (HGF) for 10 min. The cells were then analyzed with immunoblotting. (e) Viability of cells cultured in the presence of various concentrations of serum for 3 days. Bars, SD (n = 4). *P < 0.005 and **P < 0.0005 by Student's t-test. (f) 44As3 and 58As9 cells were inoculated intraperitoneally into nude mice, and DMSO (vehicle) or the Met inhibitor PHA was administered. Representative macroscopic views of mesentery tumor nodules are shown. (g) The number of mesentery tumor nodules (>1 mm in diameter). Bars, SEM (n = 11 for DMSO and 13 for PHA in 44As3; n = 10 for DMSO and PHA in 58As9). *P < 0.001 by the Mann–Whitney test.
Mentions: The IC50 values of Met inhibitors in 44As3 and 58As9 cells were approximately 1500 and 19 nM for PHA-665752, >10 000 and 6.9 nM for JNJ-38877605, and 2600 and 21 nM for PF-2341066, respectively (Fig. S2a). Met inhibitors significantly blocked phosphorylation of Met in both 44As3 and 58As9 cells (Fig.2a). Nonetheless, overall protein tyrosine phosphorylation and phosphorylation of ERK, Akt and Stat3 were significantly decreased only in 58As9 cells (Fig.2a,b). Interestingly, Src phosphorylation was rather increased in 58As9 cells upon Met inhibition, whereas it was unchanged in 44As3 cells (Fig.2a). Similar results were obtained with PF-2341066 (Fig. S2b). Met inhibitor treatment also induced marked changes in cell morphology in 58As9 but not in 44As3 cells: rounded morphology was converted to flattened and adherent phenotypes (Fig.2c). Serum starvation reduced and following HGF stimulation increased Met phosphorylation in 44As3 cells, whereas Met was constitutively phosphorylated in 58As9 cells (Fig.2d). Consequently, growth of 44As3 but not of 58As9 cells was sensitive to serum starvation (Fig.2e). These results indicate that overexpressed Met proteins confer the capability for serum-independent growth to 58As9 cells via activation of downstream signaling pathways, and, therefore, that 58As9 cells are susceptible to Met inhibitors.

Bottom Line: Saracatinib also effectively impaired peritoneal dissemination of Met-independent and FGFR2-independent SGC cells.Moreover, DGC cell lines exhibited nearly mutually exclusive susceptibility to Met, FGFR and Src inhibitors.These results suggest that DGC have distinct sensitivities to molecular target drugs and that targeting Src is beneficial in the treatment of DGC insensitive to Met and FGFR inhibition.

View Article: PubMed Central - PubMed

Affiliation: Division of Metastasis and Invasion Signaling, National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan.

Show MeSH
Related in: MedlinePlus