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Reduced Expression of Galectin-9 Contributes to a Poor Outcome in Colon Cancer by Inhibiting NK Cell Chemotaxis Partially through the Rho/ROCK1 Signaling Pathway.

Wang Y, Sun J, Ma C, Gao W, Song B, Xue H, Chen W, Chen X, Zhang Y, Shao Q, Wang Q, Zhao L, Liu J, Wang X, Wang H, Zhang Y, Yang M, Qu X - PLoS ONE (2016)

Bottom Line: A Kaplan-Meier method and Cox proportional hazards regression analysis showed that overall survival was longer in patients with high galectin-9 expression in an 8-year follow-up (P<0.05).Spearman's rank correlation indicated that there was a linear correlation between galectin-9 expression and CD56+ NK cell infiltration (R(2) = 0.658; P<0.0001).Galectin-9 stimulated migration in human NK-92 cells by affecting F-actin polarization through the Rho/ROCK1 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences, Qilu Hospital, Shandong University, Jinan, Shandong, People's Republic of China.

ABSTRACT
Galectin-9 is a widely expressed protein that is involved in immune regulation and tumorpathogenesis and serves as a marker of a poor prognosis in various types of cancers. However, the clinical impact and the precise mechanism by which this protein contributes to colon tumor progression are unclear. In the present study, we detected the expression of galectin-9 and CD56 cells using immunohistochemistry. Spearman's rank correlation was used to clarify the association between galectin-9 expression and natural killer (NK) cell infiltration. The influence of galectin-9 on NK-92 cell migration was evaluated in vitro using transwell chemotaxis assays. The role of rh-galectin-9 in F-actin polarization in NK-92 cells was investigated using laser scanning confocal microscopy. We showed that galectin-9 was expressed in 101 (78.91%) colon tumor tissues and that was expressed at lower levels in these tissues than in para-tumor tissues. Low levels of galectin-9 expression were positively correlated with a poor histological grade and lymph node metastasis (P<0.05). A Kaplan-Meier method and Cox proportional hazards regression analysis showed that overall survival was longer in patients with high galectin-9 expression in an 8-year follow-up (P<0.05). Spearman's rank correlation indicated that there was a linear correlation between galectin-9 expression and CD56+ NK cell infiltration (R(2) = 0.658; P<0.0001). Galectin-9 stimulated migration in human NK-92 cells by affecting F-actin polarization through the Rho/ROCK1 signaling pathway. These results suggest that galectin-9 expression potentially represents a novel mechanism for tumors to escape immune surveillance in colon tumors.

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Galectin-9 enhances primary NK cell chemotaxis by increasing Rho family expression.Chemotaxis of primary NK cells in response to HT29 supernatants treated with galectin-9 siRNA(A) or varying concentrations of galectin-9(B). A migration index (MI) was used to account for the high amount of random migration that occurred in primary NK cells. IL-12 was used as a positive control for initiating chemotaxis primary NK cells. * P<0.05 vs. control. And # P<0.05 vs. NC. (C), Chemotaxis was inhibited by Rho and ROCK inhibitors primary NK cells. Primary NK cells were stimulated using medium alone for 4 h or medium containing Y27632 (20 μM) or C3 transferase (2 μg/mL) for 1.5 h. * P<0.05 vs. control. (D), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (E), the expression of ROCK1 protein in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 24 h. Representative data are shown from at least 3 experiments.
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pone.0152599.g006: Galectin-9 enhances primary NK cell chemotaxis by increasing Rho family expression.Chemotaxis of primary NK cells in response to HT29 supernatants treated with galectin-9 siRNA(A) or varying concentrations of galectin-9(B). A migration index (MI) was used to account for the high amount of random migration that occurred in primary NK cells. IL-12 was used as a positive control for initiating chemotaxis primary NK cells. * P<0.05 vs. control. And # P<0.05 vs. NC. (C), Chemotaxis was inhibited by Rho and ROCK inhibitors primary NK cells. Primary NK cells were stimulated using medium alone for 4 h or medium containing Y27632 (20 μM) or C3 transferase (2 μg/mL) for 1.5 h. * P<0.05 vs. control. (D), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (E), the expression of ROCK1 protein in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 24 h. Representative data are shown from at least 3 experiments.

Mentions: In order to further validate our results, we perform the main experiments with primary NK cells. The results show that the migration of primary NK cells can be influenced by both galectin-9 secreted by HT29 (Fig 6A) or rh-galectin-9 (Fig 6B). Rho and ROCK inhibitors inhibit the chemotaxis of primary NK cell (Fig 6C). In addition, rh-galectin-9 also increase the expression of Rho family (Fig 6D) and ROCK1(Fig 6E).


Reduced Expression of Galectin-9 Contributes to a Poor Outcome in Colon Cancer by Inhibiting NK Cell Chemotaxis Partially through the Rho/ROCK1 Signaling Pathway.

Wang Y, Sun J, Ma C, Gao W, Song B, Xue H, Chen W, Chen X, Zhang Y, Shao Q, Wang Q, Zhao L, Liu J, Wang X, Wang H, Zhang Y, Yang M, Qu X - PLoS ONE (2016)

Galectin-9 enhances primary NK cell chemotaxis by increasing Rho family expression.Chemotaxis of primary NK cells in response to HT29 supernatants treated with galectin-9 siRNA(A) or varying concentrations of galectin-9(B). A migration index (MI) was used to account for the high amount of random migration that occurred in primary NK cells. IL-12 was used as a positive control for initiating chemotaxis primary NK cells. * P<0.05 vs. control. And # P<0.05 vs. NC. (C), Chemotaxis was inhibited by Rho and ROCK inhibitors primary NK cells. Primary NK cells were stimulated using medium alone for 4 h or medium containing Y27632 (20 μM) or C3 transferase (2 μg/mL) for 1.5 h. * P<0.05 vs. control. (D), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (E), the expression of ROCK1 protein in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 24 h. Representative data are shown from at least 3 experiments.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4814049&req=5

pone.0152599.g006: Galectin-9 enhances primary NK cell chemotaxis by increasing Rho family expression.Chemotaxis of primary NK cells in response to HT29 supernatants treated with galectin-9 siRNA(A) or varying concentrations of galectin-9(B). A migration index (MI) was used to account for the high amount of random migration that occurred in primary NK cells. IL-12 was used as a positive control for initiating chemotaxis primary NK cells. * P<0.05 vs. control. And # P<0.05 vs. NC. (C), Chemotaxis was inhibited by Rho and ROCK inhibitors primary NK cells. Primary NK cells were stimulated using medium alone for 4 h or medium containing Y27632 (20 μM) or C3 transferase (2 μg/mL) for 1.5 h. * P<0.05 vs. control. (D), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (E), the expression of ROCK1 protein in primary NK cells that were incubated in the presence or absence of rhGalectin-9 at a concentration of 100 ng/mL for 24 h. Representative data are shown from at least 3 experiments.
Mentions: In order to further validate our results, we perform the main experiments with primary NK cells. The results show that the migration of primary NK cells can be influenced by both galectin-9 secreted by HT29 (Fig 6A) or rh-galectin-9 (Fig 6B). Rho and ROCK inhibitors inhibit the chemotaxis of primary NK cell (Fig 6C). In addition, rh-galectin-9 also increase the expression of Rho family (Fig 6D) and ROCK1(Fig 6E).

Bottom Line: A Kaplan-Meier method and Cox proportional hazards regression analysis showed that overall survival was longer in patients with high galectin-9 expression in an 8-year follow-up (P<0.05).Spearman's rank correlation indicated that there was a linear correlation between galectin-9 expression and CD56+ NK cell infiltration (R(2) = 0.658; P<0.0001).Galectin-9 stimulated migration in human NK-92 cells by affecting F-actin polarization through the Rho/ROCK1 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Institute of Basic Medical Sciences, Qilu Hospital, Shandong University, Jinan, Shandong, People's Republic of China.

ABSTRACT
Galectin-9 is a widely expressed protein that is involved in immune regulation and tumorpathogenesis and serves as a marker of a poor prognosis in various types of cancers. However, the clinical impact and the precise mechanism by which this protein contributes to colon tumor progression are unclear. In the present study, we detected the expression of galectin-9 and CD56 cells using immunohistochemistry. Spearman's rank correlation was used to clarify the association between galectin-9 expression and natural killer (NK) cell infiltration. The influence of galectin-9 on NK-92 cell migration was evaluated in vitro using transwell chemotaxis assays. The role of rh-galectin-9 in F-actin polarization in NK-92 cells was investigated using laser scanning confocal microscopy. We showed that galectin-9 was expressed in 101 (78.91%) colon tumor tissues and that was expressed at lower levels in these tissues than in para-tumor tissues. Low levels of galectin-9 expression were positively correlated with a poor histological grade and lymph node metastasis (P<0.05). A Kaplan-Meier method and Cox proportional hazards regression analysis showed that overall survival was longer in patients with high galectin-9 expression in an 8-year follow-up (P<0.05). Spearman's rank correlation indicated that there was a linear correlation between galectin-9 expression and CD56+ NK cell infiltration (R(2) = 0.658; P<0.0001). Galectin-9 stimulated migration in human NK-92 cells by affecting F-actin polarization through the Rho/ROCK1 signaling pathway. These results suggest that galectin-9 expression potentially represents a novel mechanism for tumors to escape immune surveillance in colon tumors.

Show MeSH
Related in: MedlinePlus