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Loss of hepatocyte growth factor activator inhibitor type 1 participates in metastatic spreading of human pancreatic cancer cells in a mouse orthotopic transplantation model.

Ye J, Kawaguchi M, Haruyama Y, Kanemaru A, Fukushima T, Yamamoto K, Lin CY, Kataoka H - Cancer Sci. (2013)

Bottom Line: Previously we reported that S2-CP8 cells, a metastatic variant of the SUIT-2 human pancreatic adenocarcinoma cell line, showed markedly decreased HAI-1 expression.In vitro migration and invasion assays revealed inhibitory effects of HAI-1 on S2-CP8 cell migration and invasion.These data indicate that HAI-1 loss contributes to invasion and dissemination of a highly metastatic subline of SUIT-2, suggesting crucial roles for the balance of pericellular serine proteases/inhibitors in pancreatic cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Section of Oncopathology and Regenerative Biology, Department of Pathology, University of Miyazaki, Miyazaki, Japan; Clinical Research Center, The 2nd Affiliated Hospital School of Medicine, Zhejiang University, Hangzhou, China.

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Effect of PAR-2 KD on Matrigel invasion of S2-CP8_HAI-1tet#1 cells. (a) Effect of PAR-2 antagonist on Matrigel invasion (24 h incubation). (b) Effect of PAR-2 KD on Matrigel invasion (24 h incubation). The KD effect was verified by immunoblot and shown. Data are the mean ± SD. *P = 0.017; **P < 0.0001 (Mann–Whitney U-tests).
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fig04: Effect of PAR-2 KD on Matrigel invasion of S2-CP8_HAI-1tet#1 cells. (a) Effect of PAR-2 antagonist on Matrigel invasion (24 h incubation). (b) Effect of PAR-2 KD on Matrigel invasion (24 h incubation). The KD effect was verified by immunoblot and shown. Data are the mean ± SD. *P = 0.017; **P < 0.0001 (Mann–Whitney U-tests).

Mentions: Protease-activated receptor-2 is an epithelial cell surface protein that is activated by matriptase,16,17 and is suggested to be involved in the invasive growth of pancreatic cancer.31,32 As S2-CP8 cells express PAR-2, we hypothesized that excess PAR-2 activation may be involved in HAI-1 loss-induced enhanced migration and invasion. To test this hypothesis, we examined the effects of PAR-2 antagonist on the migration and invasion of S2-CP8_HAI-1tet#1 cells with or without Tet treatment. A selective antagonist of PAR-2 (FSLLRY) inhibited migration of S2-CP8_HAI-1tet#1 cells in the absence but not presence of Tet, which cancelled out the advantage of cellular migration conferred by loss of HAI-1 (Fig.3a). In contrast, addition of a PAR-2-activating peptide (SLIGR) significantly enhanced the migration of both HAI-1-expressing and control S2-CP8 cells. In a Matrigel invasion assay, a PAR-2 antagonist also showed a suppressive effect on the invasion activity of control S2-CP8, but not on HAI-1-expressing S2-CP8 cells (Fig.3b). While the suppression effect was significant in S2-CP8_mock and S2-CP8_HAI-1tet#2, it was modest and not statistically significant in S2-CP8/HAI-1tet#1 after 48 h incubation. However, at an earlier time point (24 h incubation), the difference was apparent (P = 0.017) in S2-CP8_HAI-1#1, and PAR-2 KD, instead of the antagonist treatment, resulted in significant suppression of the invasion of S2-CP8/HAI-1tet#1 cells in the absence of Tet (Fig.4). Taken together, these results indicate that HAI-1 loss-induced enhanced invasion of S2-CP8 cells is mediated, at least partly, by matriptase-mediated activation of PAR-2.


Loss of hepatocyte growth factor activator inhibitor type 1 participates in metastatic spreading of human pancreatic cancer cells in a mouse orthotopic transplantation model.

Ye J, Kawaguchi M, Haruyama Y, Kanemaru A, Fukushima T, Yamamoto K, Lin CY, Kataoka H - Cancer Sci. (2013)

Effect of PAR-2 KD on Matrigel invasion of S2-CP8_HAI-1tet#1 cells. (a) Effect of PAR-2 antagonist on Matrigel invasion (24 h incubation). (b) Effect of PAR-2 KD on Matrigel invasion (24 h incubation). The KD effect was verified by immunoblot and shown. Data are the mean ± SD. *P = 0.017; **P < 0.0001 (Mann–Whitney U-tests).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317873&req=5

fig04: Effect of PAR-2 KD on Matrigel invasion of S2-CP8_HAI-1tet#1 cells. (a) Effect of PAR-2 antagonist on Matrigel invasion (24 h incubation). (b) Effect of PAR-2 KD on Matrigel invasion (24 h incubation). The KD effect was verified by immunoblot and shown. Data are the mean ± SD. *P = 0.017; **P < 0.0001 (Mann–Whitney U-tests).
Mentions: Protease-activated receptor-2 is an epithelial cell surface protein that is activated by matriptase,16,17 and is suggested to be involved in the invasive growth of pancreatic cancer.31,32 As S2-CP8 cells express PAR-2, we hypothesized that excess PAR-2 activation may be involved in HAI-1 loss-induced enhanced migration and invasion. To test this hypothesis, we examined the effects of PAR-2 antagonist on the migration and invasion of S2-CP8_HAI-1tet#1 cells with or without Tet treatment. A selective antagonist of PAR-2 (FSLLRY) inhibited migration of S2-CP8_HAI-1tet#1 cells in the absence but not presence of Tet, which cancelled out the advantage of cellular migration conferred by loss of HAI-1 (Fig.3a). In contrast, addition of a PAR-2-activating peptide (SLIGR) significantly enhanced the migration of both HAI-1-expressing and control S2-CP8 cells. In a Matrigel invasion assay, a PAR-2 antagonist also showed a suppressive effect on the invasion activity of control S2-CP8, but not on HAI-1-expressing S2-CP8 cells (Fig.3b). While the suppression effect was significant in S2-CP8_mock and S2-CP8_HAI-1tet#2, it was modest and not statistically significant in S2-CP8/HAI-1tet#1 after 48 h incubation. However, at an earlier time point (24 h incubation), the difference was apparent (P = 0.017) in S2-CP8_HAI-1#1, and PAR-2 KD, instead of the antagonist treatment, resulted in significant suppression of the invasion of S2-CP8/HAI-1tet#1 cells in the absence of Tet (Fig.4). Taken together, these results indicate that HAI-1 loss-induced enhanced invasion of S2-CP8 cells is mediated, at least partly, by matriptase-mediated activation of PAR-2.

Bottom Line: Previously we reported that S2-CP8 cells, a metastatic variant of the SUIT-2 human pancreatic adenocarcinoma cell line, showed markedly decreased HAI-1 expression.In vitro migration and invasion assays revealed inhibitory effects of HAI-1 on S2-CP8 cell migration and invasion.These data indicate that HAI-1 loss contributes to invasion and dissemination of a highly metastatic subline of SUIT-2, suggesting crucial roles for the balance of pericellular serine proteases/inhibitors in pancreatic cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Section of Oncopathology and Regenerative Biology, Department of Pathology, University of Miyazaki, Miyazaki, Japan; Clinical Research Center, The 2nd Affiliated Hospital School of Medicine, Zhejiang University, Hangzhou, China.

Show MeSH
Related in: MedlinePlus