Limits...
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.

Show MeSH

Related in: MedlinePlus

Reoxygenation enhances the malignant potential of PDAC cells. (a) Representative images of CA9 and HIF-1α expression by immunofluorescence in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells derived from AsPC-1 (left panels) (×400). The graph shows the relative ratio of CA9 or HIF-1α positive cells (right panels). (b) Cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were evaluated (n = 3). (c) Invaded cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were analyzed (n = 3). The graph shows mean ± SD. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317936&req=5

fig01: Reoxygenation enhances the malignant potential of PDAC cells. (a) Representative images of CA9 and HIF-1α expression by immunofluorescence in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells derived from AsPC-1 (left panels) (×400). The graph shows the relative ratio of CA9 or HIF-1α positive cells (right panels). (b) Cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were evaluated (n = 3). (c) Invaded cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were analyzed (n = 3). The graph shows mean ± SD. *P < 0.05.

Mentions: Wild type cells, chronic hypoxia-resistant PDAC cells (Ch-H-R cells), PDAC cells cultured under acute hypoxia (Ac-H cells) and reoxygenated Ch-H-R cells were used to investigate the change of phenotypes by reoxygenation. First, we evaluated HIF-1α, well known as one of the main hypoxic transcriptional factors, and its downstream protein CA9. CA9 positive cells were counted as described in Fig. S3. CA9 and HIF-1α were expressed in Ch-H-R cells and Ac-H cells. However, the amount of CA9 or HIF-1α positive cells in reoxygenated Ch-H-R cells was significantly lower than in Ch-H-R cells and Ac-H cells (Figs1a,S4), and CA9 and HIF-1α protein expression were also confirmed by western blotting analysis (Fig. S5). Next, we examined proliferation and invasiveness, factors of malignant potential. The ability of anchorage-dependent proliferation of reoxygenated Ch-H-R cells was the highest among the four types of cells both in AsPC-1 and SUIT-2 (Fig.1b). Invasiveness in Ch-H-R cells was significantly higher than that in Ac-H cells, and also significantly higher in reoxygenated Ch-H-R cells than in Ch-H-R cells both in AsPC-1 and SUIT-2 (Fig.1c). These results suggest that reoxygenation confers a more aggressive phenotype in PDAC cells.


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)

Reoxygenation enhances the malignant potential of PDAC cells. (a) Representative images of CA9 and HIF-1α expression by immunofluorescence in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells derived from AsPC-1 (left panels) (×400). The graph shows the relative ratio of CA9 or HIF-1α positive cells (right panels). (b) Cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were evaluated (n = 3). (c) Invaded cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were analyzed (n = 3). The graph shows mean ± SD. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: Reoxygenation enhances the malignant potential of PDAC cells. (a) Representative images of CA9 and HIF-1α expression by immunofluorescence in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells derived from AsPC-1 (left panels) (×400). The graph shows the relative ratio of CA9 or HIF-1α positive cells (right panels). (b) Cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were evaluated (n = 3). (c) Invaded cell number in wild type cells, Ac-H cells, Ch-H-R cells and reoxygenated Ch-H-R cells were analyzed (n = 3). The graph shows mean ± SD. *P < 0.05.
Mentions: Wild type cells, chronic hypoxia-resistant PDAC cells (Ch-H-R cells), PDAC cells cultured under acute hypoxia (Ac-H cells) and reoxygenated Ch-H-R cells were used to investigate the change of phenotypes by reoxygenation. First, we evaluated HIF-1α, well known as one of the main hypoxic transcriptional factors, and its downstream protein CA9. CA9 positive cells were counted as described in Fig. S3. CA9 and HIF-1α were expressed in Ch-H-R cells and Ac-H cells. However, the amount of CA9 or HIF-1α positive cells in reoxygenated Ch-H-R cells was significantly lower than in Ch-H-R cells and Ac-H cells (Figs1a,S4), and CA9 and HIF-1α protein expression were also confirmed by western blotting analysis (Fig. S5). Next, we examined proliferation and invasiveness, factors of malignant potential. The ability of anchorage-dependent proliferation of reoxygenated Ch-H-R cells was the highest among the four types of cells both in AsPC-1 and SUIT-2 (Fig.1b). Invasiveness in Ch-H-R cells was significantly higher than that in Ac-H cells, and also significantly higher in reoxygenated Ch-H-R cells than in Ch-H-R cells both in AsPC-1 and SUIT-2 (Fig.1c). These results suggest that reoxygenation confers a more aggressive phenotype in PDAC cells.

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