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Hedgehog signaling pathway is a potential therapeutic target for gallbladder cancer.

Matsushita S, Onishi H, Nakano K, Nagamatsu I, Imaizumi A, Hattori M, Oda Y, Tanaka M, Katano M - Cancer Sci. (2014)

Bottom Line: New effective therapeutic strategies are greatly needed.In contrast, inhibiting the effector Smo decreased the anchor-dependent and anchor-independent proliferation.These results suggest that Hh signaling is elevated in GBC and may be involved in the acquisition of malignant phenotypes, and that Hh signaling may be a potential therapeutic target for GBC.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.

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Hedgehog (Hh) signaling enhances gallbladder cancer colony formation in vitro and inhibition of Smoothened (Smo) suppresses tumor growth in vivo. (a, b) GBd15 and TGBC2TKB cells were incubated with recombinant human Sonic Hh (rhShh) (a, at 0.1 or 1.0 μg/mL) or cyclopamine (Cyclo) (b, at 5 or 10 μM) for 14 days before colony formation was assessed. (c) GBd15 and TGBC2TKB cells were transfected with Smo siRNA (siSmo) and incubated for 14 days before colony formation was assessed. (d, e) TGBC2TKB cells were transfected with control siRNA (siCont) (n = 5) or Smo siRNA (n = 5), and then implanted into the flanks of athymic nude mice. The data represent the volume of each tumor (red arrows). Student's t-test was carried out between the tumor volumes of the two groups. (f) Gli1 and Smo expression were confirmed by immunohistochemistry in tumor tissue explants from the nude mice. Original magnification, ×400. Error bars represent standard deviations. *P < 0.05.
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fig07: Hedgehog (Hh) signaling enhances gallbladder cancer colony formation in vitro and inhibition of Smoothened (Smo) suppresses tumor growth in vivo. (a, b) GBd15 and TGBC2TKB cells were incubated with recombinant human Sonic Hh (rhShh) (a, at 0.1 or 1.0 μg/mL) or cyclopamine (Cyclo) (b, at 5 or 10 μM) for 14 days before colony formation was assessed. (c) GBd15 and TGBC2TKB cells were transfected with Smo siRNA (siSmo) and incubated for 14 days before colony formation was assessed. (d, e) TGBC2TKB cells were transfected with control siRNA (siCont) (n = 5) or Smo siRNA (n = 5), and then implanted into the flanks of athymic nude mice. The data represent the volume of each tumor (red arrows). Student's t-test was carried out between the tumor volumes of the two groups. (f) Gli1 and Smo expression were confirmed by immunohistochemistry in tumor tissue explants from the nude mice. Original magnification, ×400. Error bars represent standard deviations. *P < 0.05.

Mentions: We next investigated the role of Hh signaling on anchorage-independent growth of GBC cells. The addition of rhShh significantly enhanced colony formation, an anchorage-independent phenotype, in GBd15 and TGBC2TKB cells (Fig.7a). Cyclopamine and Smo siRNA also significantly suppressed colony formation in GBd15 and TGBC2TKB cells (Fig.7b,c). These results indicate that Hh signaling affects anchorage-independent growth in GBC cells. To determine whether the observed changes induced by Hh signaling in cells is reflected in vivo, we investigated the tumorigenicity of GBC cells transfected with Smo siRNA in athymic nude mice. Subcutaneous tumors developed in three out of five mice injected with cells transfected with Smo siRNA. In contrast, all of the five mice injected with control siRNA transfected cells showed tumor development (Fig.7d). There were significant differences in tumor growth/size between Smo siRNA and control siRNA groups as well (Fig.7e). Furthermore, we confirmed that the expression of Gli1 and Smo in Smo siRNA transfected GBC cells were lower than in the controls (Fig.7f).


Hedgehog signaling pathway is a potential therapeutic target for gallbladder cancer.

Matsushita S, Onishi H, Nakano K, Nagamatsu I, Imaizumi A, Hattori M, Oda Y, Tanaka M, Katano M - Cancer Sci. (2014)

Hedgehog (Hh) signaling enhances gallbladder cancer colony formation in vitro and inhibition of Smoothened (Smo) suppresses tumor growth in vivo. (a, b) GBd15 and TGBC2TKB cells were incubated with recombinant human Sonic Hh (rhShh) (a, at 0.1 or 1.0 μg/mL) or cyclopamine (Cyclo) (b, at 5 or 10 μM) for 14 days before colony formation was assessed. (c) GBd15 and TGBC2TKB cells were transfected with Smo siRNA (siSmo) and incubated for 14 days before colony formation was assessed. (d, e) TGBC2TKB cells were transfected with control siRNA (siCont) (n = 5) or Smo siRNA (n = 5), and then implanted into the flanks of athymic nude mice. The data represent the volume of each tumor (red arrows). Student's t-test was carried out between the tumor volumes of the two groups. (f) Gli1 and Smo expression were confirmed by immunohistochemistry in tumor tissue explants from the nude mice. Original magnification, ×400. Error bars represent standard deviations. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317941&req=5

fig07: Hedgehog (Hh) signaling enhances gallbladder cancer colony formation in vitro and inhibition of Smoothened (Smo) suppresses tumor growth in vivo. (a, b) GBd15 and TGBC2TKB cells were incubated with recombinant human Sonic Hh (rhShh) (a, at 0.1 or 1.0 μg/mL) or cyclopamine (Cyclo) (b, at 5 or 10 μM) for 14 days before colony formation was assessed. (c) GBd15 and TGBC2TKB cells were transfected with Smo siRNA (siSmo) and incubated for 14 days before colony formation was assessed. (d, e) TGBC2TKB cells were transfected with control siRNA (siCont) (n = 5) or Smo siRNA (n = 5), and then implanted into the flanks of athymic nude mice. The data represent the volume of each tumor (red arrows). Student's t-test was carried out between the tumor volumes of the two groups. (f) Gli1 and Smo expression were confirmed by immunohistochemistry in tumor tissue explants from the nude mice. Original magnification, ×400. Error bars represent standard deviations. *P < 0.05.
Mentions: We next investigated the role of Hh signaling on anchorage-independent growth of GBC cells. The addition of rhShh significantly enhanced colony formation, an anchorage-independent phenotype, in GBd15 and TGBC2TKB cells (Fig.7a). Cyclopamine and Smo siRNA also significantly suppressed colony formation in GBd15 and TGBC2TKB cells (Fig.7b,c). These results indicate that Hh signaling affects anchorage-independent growth in GBC cells. To determine whether the observed changes induced by Hh signaling in cells is reflected in vivo, we investigated the tumorigenicity of GBC cells transfected with Smo siRNA in athymic nude mice. Subcutaneous tumors developed in three out of five mice injected with cells transfected with Smo siRNA. In contrast, all of the five mice injected with control siRNA transfected cells showed tumor development (Fig.7d). There were significant differences in tumor growth/size between Smo siRNA and control siRNA groups as well (Fig.7e). Furthermore, we confirmed that the expression of Gli1 and Smo in Smo siRNA transfected GBC cells were lower than in the controls (Fig.7f).

Bottom Line: New effective therapeutic strategies are greatly needed.In contrast, inhibiting the effector Smo decreased the anchor-dependent and anchor-independent proliferation.These results suggest that Hh signaling is elevated in GBC and may be involved in the acquisition of malignant phenotypes, and that Hh signaling may be a potential therapeutic target for GBC.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.

Show MeSH
Related in: MedlinePlus