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Macrophage Infiltration Induces Gastric Cancer Invasiveness by Activating the β-Catenin Pathway.

Wu MH, Lee WJ, Hua KT, Kuo ML, Lin MT - PLoS ONE (2015)

Bottom Line: Increased density of macrophages was associated with advanced stage and poor survival.AKT but not ERK regulated β-catenin translocation.Macrophages may induce invasiveness by activating the β-catenin pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan.

ABSTRACT

Background: Despite evidence that activated macrophages act in an inflammatory microenvironment to promote gastric tumorigenesis via β-catenin signaling, the effects of β-catenin signaling on gastric cancer cell metastasis and the relationship of these cells with surrounding tumor associated macrophages have not been directly studied.

Methods: Immunohistochemical staining was employed to analyze 103 patients. An invasion assay was used to evaluate the relationship between macrophages and gastric cancer cells. β-catenin gain-of-function and loss-of-function approaches were performed. To assess the β-catenin regulation mechanism in gastric cancer cells, Western blotting and reverse-transcription polymerase chain reaction were used.

Results: Increased density of macrophages was associated with advanced stage and poor survival. Gastric cancer cell lines co-cultured with macrophages conditioned medium showed increased nuclear accumulation of β-catenin and increased invading ability. AKT but not ERK regulated β-catenin translocation. MMP7 and CD44, both β-catenin downstream genes, were involved in macrophage-activated gastric cancer cell invasion.

Conclusion(s): Collectively, the clinical data suggest that macrophage infiltration is correlated with increased grade and poor prognosis for gastric cancer patients who underwent radical resection. Macrophages may induce invasiveness by activating the β-catenin pathway.

No MeSH data available.


Related in: MedlinePlus

(A) Migration activity of THP-1 monocyte co-cultured with different gastric cancer cell lines (AGS, N87, MKN-45 and TSGH). THP-1 monocyte will be recruited by different gastric cancer cell lines and the recruiting ability is dependent on the degree of malignancy. (B) Invasion ability of GC cells treated by macrophage CM. Four different types of GC cells (AGS, MKN45, N87 and TSGH) were seeded in a Boyden chamber and co-cultured with or without macrophage cells or macrophage CM for 24 hours. Invasion abilities of each cell line were measured in vitro for 24 hours. All data represent the arithmetic mean ± SEM. * p < 0.05, ** p < 0.01. (C) Effect of macrophage CM co-cultured with cells. N87 and AGS cells were co-cultured with macrophage CM respectively for 24 hours and cell viability was determined by MTT assay.
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pone.0134122.g004: (A) Migration activity of THP-1 monocyte co-cultured with different gastric cancer cell lines (AGS, N87, MKN-45 and TSGH). THP-1 monocyte will be recruited by different gastric cancer cell lines and the recruiting ability is dependent on the degree of malignancy. (B) Invasion ability of GC cells treated by macrophage CM. Four different types of GC cells (AGS, MKN45, N87 and TSGH) were seeded in a Boyden chamber and co-cultured with or without macrophage cells or macrophage CM for 24 hours. Invasion abilities of each cell line were measured in vitro for 24 hours. All data represent the arithmetic mean ± SEM. * p < 0.05, ** p < 0.01. (C) Effect of macrophage CM co-cultured with cells. N87 and AGS cells were co-cultured with macrophage CM respectively for 24 hours and cell viability was determined by MTT assay.

Mentions: To further confirm our clinical findings in vitro, we first co-cultured macrophages with different GC cell lines (AGS, N87, MKN-45 and TSGH). All GC cell lines tested would recruit macrophages to migrate and the macrophage-recruiting ability was dependent on the degree of malignancy of the cancer cells as determined by their invasiveness (Fig 4A).[11] To evaluate whether invasiveness or proliferation of GC cells can be regulated by macrophages, AGS, N87, MKN45 and TSGH were co-cultured with and without macrophages or CM from macrophage for 24 h and their invasion and proliferation abilities tested. All cell types co-cultured with macrophage or CM from macrophage showed an near 2-fold increase in the number of invading cells (compared with negative controls, p < 0.05), which is not associated with their proliferation ability (Fig 4B–4D).


Macrophage Infiltration Induces Gastric Cancer Invasiveness by Activating the β-Catenin Pathway.

Wu MH, Lee WJ, Hua KT, Kuo ML, Lin MT - PLoS ONE (2015)

(A) Migration activity of THP-1 monocyte co-cultured with different gastric cancer cell lines (AGS, N87, MKN-45 and TSGH). THP-1 monocyte will be recruited by different gastric cancer cell lines and the recruiting ability is dependent on the degree of malignancy. (B) Invasion ability of GC cells treated by macrophage CM. Four different types of GC cells (AGS, MKN45, N87 and TSGH) were seeded in a Boyden chamber and co-cultured with or without macrophage cells or macrophage CM for 24 hours. Invasion abilities of each cell line were measured in vitro for 24 hours. All data represent the arithmetic mean ± SEM. * p < 0.05, ** p < 0.01. (C) Effect of macrophage CM co-cultured with cells. N87 and AGS cells were co-cultured with macrophage CM respectively for 24 hours and cell viability was determined by MTT assay.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520459&req=5

pone.0134122.g004: (A) Migration activity of THP-1 monocyte co-cultured with different gastric cancer cell lines (AGS, N87, MKN-45 and TSGH). THP-1 monocyte will be recruited by different gastric cancer cell lines and the recruiting ability is dependent on the degree of malignancy. (B) Invasion ability of GC cells treated by macrophage CM. Four different types of GC cells (AGS, MKN45, N87 and TSGH) were seeded in a Boyden chamber and co-cultured with or without macrophage cells or macrophage CM for 24 hours. Invasion abilities of each cell line were measured in vitro for 24 hours. All data represent the arithmetic mean ± SEM. * p < 0.05, ** p < 0.01. (C) Effect of macrophage CM co-cultured with cells. N87 and AGS cells were co-cultured with macrophage CM respectively for 24 hours and cell viability was determined by MTT assay.
Mentions: To further confirm our clinical findings in vitro, we first co-cultured macrophages with different GC cell lines (AGS, N87, MKN-45 and TSGH). All GC cell lines tested would recruit macrophages to migrate and the macrophage-recruiting ability was dependent on the degree of malignancy of the cancer cells as determined by their invasiveness (Fig 4A).[11] To evaluate whether invasiveness or proliferation of GC cells can be regulated by macrophages, AGS, N87, MKN45 and TSGH were co-cultured with and without macrophages or CM from macrophage for 24 h and their invasion and proliferation abilities tested. All cell types co-cultured with macrophage or CM from macrophage showed an near 2-fold increase in the number of invading cells (compared with negative controls, p < 0.05), which is not associated with their proliferation ability (Fig 4B–4D).

Bottom Line: Increased density of macrophages was associated with advanced stage and poor survival.AKT but not ERK regulated β-catenin translocation.Macrophages may induce invasiveness by activating the β-catenin pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan.

ABSTRACT

Background: Despite evidence that activated macrophages act in an inflammatory microenvironment to promote gastric tumorigenesis via β-catenin signaling, the effects of β-catenin signaling on gastric cancer cell metastasis and the relationship of these cells with surrounding tumor associated macrophages have not been directly studied.

Methods: Immunohistochemical staining was employed to analyze 103 patients. An invasion assay was used to evaluate the relationship between macrophages and gastric cancer cells. β-catenin gain-of-function and loss-of-function approaches were performed. To assess the β-catenin regulation mechanism in gastric cancer cells, Western blotting and reverse-transcription polymerase chain reaction were used.

Results: Increased density of macrophages was associated with advanced stage and poor survival. Gastric cancer cell lines co-cultured with macrophages conditioned medium showed increased nuclear accumulation of β-catenin and increased invading ability. AKT but not ERK regulated β-catenin translocation. MMP7 and CD44, both β-catenin downstream genes, were involved in macrophage-activated gastric cancer cell invasion.

Conclusion(s): Collectively, the clinical data suggest that macrophage infiltration is correlated with increased grade and poor prognosis for gastric cancer patients who underwent radical resection. Macrophages may induce invasiveness by activating the β-catenin pathway.

No MeSH data available.


Related in: MedlinePlus