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Reoxygenation from chronic hypoxia promotes metastatic processes in pancreatic cancer through the Hedgehog signaling.

Morifuji Y, Onishi H, Iwasaki H, Imaizumi A, Nakano K, Tanaka M, Katano M - Cancer Sci. (2014)

Bottom Line: A Hedgehog (Hh) signaling component, Gli1, was significantly increased by reoxygenation.Gli1 knockdown inhibited reoxygenation-induced increases in proliferation and tumorigenicity and decreased invasiveness through suppression of matrix metalloproteinase (MMP) 2 and MMP9.These results suggest that metastatic processes in PDAC are induced through activation of the Hh signaling pathway.

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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Gli1 contributes to the augmentation of malignant potential by reoxygenation. (a) Cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was counted (n = 3). (b) Invaded cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was investigated (n = 3). (c) Reoxygenated Ch-H-R cells were transfected with control, MMP2 or MMP9 siRNA. Then the invaded cell number in these cells was counted (n = 3). (d) MMP2 and MMP9 mRNA expression in Gli1 siRNA transfected-reoxygenated Ch-H-R cells were evaluated by real-time RT-PCR (n = 3). The graph shows mean ± SD. *P < 0.05.
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fig03: Gli1 contributes to the augmentation of malignant potential by reoxygenation. (a) Cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was counted (n = 3). (b) Invaded cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was investigated (n = 3). (c) Reoxygenated Ch-H-R cells were transfected with control, MMP2 or MMP9 siRNA. Then the invaded cell number in these cells was counted (n = 3). (d) MMP2 and MMP9 mRNA expression in Gli1 siRNA transfected-reoxygenated Ch-H-R cells were evaluated by real-time RT-PCR (n = 3). The graph shows mean ± SD. *P < 0.05.

Mentions: Next, the correlation between activation of Hh signaling and reoxygenation-induced proliferation and invasiveness was investigated. Transfection with Gli1 siRNA in reoxygenated Ch-H-R cells led to an 80–90% decrease in Gli1 mRNA expression (Fig. S10). Gli1 siRNA-transfected reoxygenated cells showed significant decreases in anchorage-dependent proliferation and invasion (Fig.3a,b). To investigate the effect of MMP on invasiveness, MMP siRNA was used. MMP2 and MMP9 knockdown efficiencies were approximately 70–90 and 60–80%, respectively (Fig. S11). The invasiveness of MMP2 and MMP9 siRNA-transfected reoxygenated Ch-H-R cells was significantly lower than that of the control cells (Fig.3c). MMP2 and MMP9 co-knockdown significantly inhibited invasion compared with MMP2 or MMP9 knockdown alone (Fig.3c). In addition, Gli1 knockdown significantly decreased the expression of MMP2 and MMP9 (Fig.3d). Next, to investigate whether the upregulation of Gli1 was through Hh signaling, Shh and Smo knockdown experiments were performed. Shh and Smo knockdown efficiencies were over 90% (Fig. S12). Shh siRNA and Smo siRNA transfected-reoxygenated Ch-H-R cells showed lower anchorage-dependent proliferative (Fig.4a,b) and invasive (Fig.4c,d) abilities than the control in both cell lines. These results suggest that enhanced cell proliferation and invasion by reoxygenation are induced through the activation of Hh signaling.


Reoxygenation from chronic hypoxia promotes metastatic processes in pancreatic cancer through the Hedgehog signaling.

Morifuji Y, Onishi H, Iwasaki H, Imaizumi A, Nakano K, Tanaka M, Katano M - Cancer Sci. (2014)

Gli1 contributes to the augmentation of malignant potential by reoxygenation. (a) Cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was counted (n = 3). (b) Invaded cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was investigated (n = 3). (c) Reoxygenated Ch-H-R cells were transfected with control, MMP2 or MMP9 siRNA. Then the invaded cell number in these cells was counted (n = 3). (d) MMP2 and MMP9 mRNA expression in Gli1 siRNA transfected-reoxygenated Ch-H-R cells were evaluated by real-time RT-PCR (n = 3). The graph shows mean ± SD. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4317936&req=5

fig03: Gli1 contributes to the augmentation of malignant potential by reoxygenation. (a) Cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was counted (n = 3). (b) Invaded cell number in control or Gli1 siRNA-transfected-reoxygenated Ch-H-R cells was investigated (n = 3). (c) Reoxygenated Ch-H-R cells were transfected with control, MMP2 or MMP9 siRNA. Then the invaded cell number in these cells was counted (n = 3). (d) MMP2 and MMP9 mRNA expression in Gli1 siRNA transfected-reoxygenated Ch-H-R cells were evaluated by real-time RT-PCR (n = 3). The graph shows mean ± SD. *P < 0.05.
Mentions: Next, the correlation between activation of Hh signaling and reoxygenation-induced proliferation and invasiveness was investigated. Transfection with Gli1 siRNA in reoxygenated Ch-H-R cells led to an 80–90% decrease in Gli1 mRNA expression (Fig. S10). Gli1 siRNA-transfected reoxygenated cells showed significant decreases in anchorage-dependent proliferation and invasion (Fig.3a,b). To investigate the effect of MMP on invasiveness, MMP siRNA was used. MMP2 and MMP9 knockdown efficiencies were approximately 70–90 and 60–80%, respectively (Fig. S11). The invasiveness of MMP2 and MMP9 siRNA-transfected reoxygenated Ch-H-R cells was significantly lower than that of the control cells (Fig.3c). MMP2 and MMP9 co-knockdown significantly inhibited invasion compared with MMP2 or MMP9 knockdown alone (Fig.3c). In addition, Gli1 knockdown significantly decreased the expression of MMP2 and MMP9 (Fig.3d). Next, to investigate whether the upregulation of Gli1 was through Hh signaling, Shh and Smo knockdown experiments were performed. Shh and Smo knockdown efficiencies were over 90% (Fig. S12). Shh siRNA and Smo siRNA transfected-reoxygenated Ch-H-R cells showed lower anchorage-dependent proliferative (Fig.4a,b) and invasive (Fig.4c,d) abilities than the control in both cell lines. These results suggest that enhanced cell proliferation and invasion by reoxygenation are induced through the activation of Hh signaling.

Bottom Line: A Hedgehog (Hh) signaling component, Gli1, was significantly increased by reoxygenation.Gli1 knockdown inhibited reoxygenation-induced increases in proliferation and tumorigenicity and decreased invasiveness through suppression of matrix metalloproteinase (MMP) 2 and MMP9.These results suggest that metastatic processes in PDAC are induced through activation of the Hh signaling pathway.

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