Limits...
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.

Show MeSH

Related in: MedlinePlus

Galectin-9 promotes F-actin rearrangement by enhancing the expression of Rho family members and ROCK1.(A), Chemotaxis was inhibited by Rho and ROCK inhibitors in NK-92 cells. NK-92 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. A migration index (MI) was used to account for the high random migration that was observed in NK cells. * P<0.05 vs. control. (B), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (C), The expression of ROCK1 protein in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 24 h. (D), Representative micrographs of NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h. F-actin was stained with rhodamine-labeled phalloidin (red), and cell nuclei were labeled with DAPI (blue) (630×). Representative data are shown from at least 3 experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4814049&req=5

pone.0152599.g005: Galectin-9 promotes F-actin rearrangement by enhancing the expression of Rho family members and ROCK1.(A), Chemotaxis was inhibited by Rho and ROCK inhibitors in NK-92 cells. NK-92 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. A migration index (MI) was used to account for the high random migration that was observed in NK cells. * P<0.05 vs. control. (B), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (C), The expression of ROCK1 protein in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 24 h. (D), Representative micrographs of NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h. F-actin was stained with rhodamine-labeled phalloidin (red), and cell nuclei were labeled with DAPI (blue) (630×). Representative data are shown from at least 3 experiments.

Mentions: Lymphocytes undergo dramatic cytoskeletal rearrangements during cell migration.The small GTPase Rho is a pivotal regulator of the actin cytoskeleton, and Rho kinase, which is also known as Rho-associated coiled coil kinase (ROCK), is a Rho effector protein. To assess the involvement of Rho/ROCK in the transmigration of NK cells, chemotaxis assays were performed in the presence of the well-characterized ROCK inhibitor, Y27632, and another Rho-specific inhibitor, C3 transferase. The results showed that Y27632 and C3 transferase significantly inhibited NK-92 cell migration (Fig 5A).


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 promotes F-actin rearrangement by enhancing the expression of Rho family members and ROCK1.(A), Chemotaxis was inhibited by Rho and ROCK inhibitors in NK-92 cells. NK-92 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. A migration index (MI) was used to account for the high random migration that was observed in NK cells. * P<0.05 vs. control. (B), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (C), The expression of ROCK1 protein in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 24 h. (D), Representative micrographs of NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h. F-actin was stained with rhodamine-labeled phalloidin (red), and cell nuclei were labeled with DAPI (blue) (630×). Representative data are shown from at least 3 experiments.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0152599.g005: Galectin-9 promotes F-actin rearrangement by enhancing the expression of Rho family members and ROCK1.(A), Chemotaxis was inhibited by Rho and ROCK inhibitors in NK-92 cells. NK-92 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. A migration index (MI) was used to account for the high random migration that was observed in NK cells. * P<0.05 vs. control. (B), The expression of RhoA, RhoB, RhoC, RhoF, RhoG, RhoH, RhoQ and RhoT1 mRNA using RT-PCR in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h; * P<0.05 vs. control. (C), The expression of ROCK1 protein in NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 24 h. (D), Representative micrographs of NK-92 cells that were incubated in the presence or absence of rh-galectin-9 at a concentration of 100 ng/mL for 4 h. F-actin was stained with rhodamine-labeled phalloidin (red), and cell nuclei were labeled with DAPI (blue) (630×). Representative data are shown from at least 3 experiments.
Mentions: Lymphocytes undergo dramatic cytoskeletal rearrangements during cell migration.The small GTPase Rho is a pivotal regulator of the actin cytoskeleton, and Rho kinase, which is also known as Rho-associated coiled coil kinase (ROCK), is a Rho effector protein. To assess the involvement of Rho/ROCK in the transmigration of NK cells, chemotaxis assays were performed in the presence of the well-characterized ROCK inhibitor, Y27632, and another Rho-specific inhibitor, C3 transferase. The results showed that Y27632 and C3 transferase significantly inhibited NK-92 cell migration (Fig 5A).

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