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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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Smoothened (Smo)-regulated invasion is mediated through epithelial–mesenchymal transition. (a) Quantitative RT-PCR (b) and Western blotting of E-cadherin and vimentin in Smo-siRNA transfected GBd15 and TGBC2TKB cells. (c) GBd15 and TGBC2TKB cells transfected with control siRNA (siCont) or Smo siRNA (siSmo) were immunostained with E-cadherin antibody (green) and DAPI (blue). Red arrows, E-cadherin accumulated in the membrane of Smo siRNA transfected cells. Original magnification, ×200. (d) GBd15 and TGBC2TKB cells transfected with Smo siRNA for 48 h before the number of spindle-shaped cells were counted using bright-field microscopy. Error bars represent standard deviations. *P < 0.05.
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fig06: Smoothened (Smo)-regulated invasion is mediated through epithelial–mesenchymal transition. (a) Quantitative RT-PCR (b) and Western blotting of E-cadherin and vimentin in Smo-siRNA transfected GBd15 and TGBC2TKB cells. (c) GBd15 and TGBC2TKB cells transfected with control siRNA (siCont) or Smo siRNA (siSmo) were immunostained with E-cadherin antibody (green) and DAPI (blue). Red arrows, E-cadherin accumulated in the membrane of Smo siRNA transfected cells. Original magnification, ×200. (d) GBd15 and TGBC2TKB cells transfected with Smo siRNA for 48 h before the number of spindle-shaped cells were counted using bright-field microscopy. Error bars represent standard deviations. *P < 0.05.

Mentions: Epithelial–mesenchymal transition (EMT) is another important factor in invasiveness.21 Therefore, we also examined the effect of Smo knockdown on EMT in GBd15 and TGBC2TKB cells. An increase in the expression of E-cadherin and a decrease in the expression of vimentin were seen by quantitative RT-PCR (Fig.6a) and Western blotting (Fig.6b) after Smo siRNA treatment in GBd15 and TGBC2TKB cells. Furthermore, E-cadherin accumulated in the membrane of Smo siRNA transfected cells (Fig.6c, red arrow). As shown in Figure6(d), Smo siRNA transfection also substantially reduced the percentage of spindle-shaped cells and the cells tended to aggregate. These results suggest that decreased cell invasion following Smo knockdown is because of a lack of EMT in GBCs.


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)

Smoothened (Smo)-regulated invasion is mediated through epithelial–mesenchymal transition. (a) Quantitative RT-PCR (b) and Western blotting of E-cadherin and vimentin in Smo-siRNA transfected GBd15 and TGBC2TKB cells. (c) GBd15 and TGBC2TKB cells transfected with control siRNA (siCont) or Smo siRNA (siSmo) were immunostained with E-cadherin antibody (green) and DAPI (blue). Red arrows, E-cadherin accumulated in the membrane of Smo siRNA transfected cells. Original magnification, ×200. (d) GBd15 and TGBC2TKB cells transfected with Smo siRNA for 48 h before the number of spindle-shaped cells were counted using bright-field microscopy. Error bars represent standard deviations. *P < 0.05.
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Related In: Results  -  Collection

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fig06: Smoothened (Smo)-regulated invasion is mediated through epithelial–mesenchymal transition. (a) Quantitative RT-PCR (b) and Western blotting of E-cadherin and vimentin in Smo-siRNA transfected GBd15 and TGBC2TKB cells. (c) GBd15 and TGBC2TKB cells transfected with control siRNA (siCont) or Smo siRNA (siSmo) were immunostained with E-cadherin antibody (green) and DAPI (blue). Red arrows, E-cadherin accumulated in the membrane of Smo siRNA transfected cells. Original magnification, ×200. (d) GBd15 and TGBC2TKB cells transfected with Smo siRNA for 48 h before the number of spindle-shaped cells were counted using bright-field microscopy. Error bars represent standard deviations. *P < 0.05.
Mentions: Epithelial–mesenchymal transition (EMT) is another important factor in invasiveness.21 Therefore, we also examined the effect of Smo knockdown on EMT in GBd15 and TGBC2TKB cells. An increase in the expression of E-cadherin and a decrease in the expression of vimentin were seen by quantitative RT-PCR (Fig.6a) and Western blotting (Fig.6b) after Smo siRNA treatment in GBd15 and TGBC2TKB cells. Furthermore, E-cadherin accumulated in the membrane of Smo siRNA transfected cells (Fig.6c, red arrow). As shown in Figure6(d), Smo siRNA transfection also substantially reduced the percentage of spindle-shaped cells and the cells tended to aggregate. These results suggest that decreased cell invasion following Smo knockdown is because of a lack of EMT in GBCs.

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