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Macrophage subtype predicts lymph node metastasis in oesophageal adenocarcinoma and promotes cancer cell invasion in vitro

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Currently, there is a lack of ideal biomarkers for predicting nodal status in preoperative stage of oesophageal adenocarcinoma (EAC) to aid optimising therapeutic options. We studied the potential of applying subtype macrophages to predict lymph node metastasis and prognosis in EAC.

Material and methods:: Fifty-three EAC resection specimens were immunostained with CD68, CD40 (M1), and CD163 (M2). Lymphatic vessel density (LVD) was estimated with the staining of D2-40. Subsequently, we tested if M2d macrophage could promote EAC cell migration and invasion.

Results:: In EAC without neoadjuvant treatment, an increase in M2-like macrophage was associated with poor patient survival, independent of the locations of macrophages in tumour. The M2/M1 ratio that represented the balance between M2- and M1-like macrophages was significantly higher in nodal-positive EACs than that in nodal-negative EACs, and inversely correlated with patient overall survival. The M2/M1 ratio was not related to LVD. EAC cell polarised THP1 cell into M2d-like macrophage, which promoted EAC cell migration and invasion. Neoadjuvant therapy appeared to diminish the correlation between the M2/M1 ratio and survival.

Conclusions:: The ratio of M2/M1 macrophage may serve as a sensitive marker to predict lymph node metastasis and poor prognosis in EAC without neoadjuvant therapy. M2d macrophage may have important roles in EAC metastasis.

No MeSH data available.


Related in: MedlinePlus

Induction of THP1 monocyte into M2-like macrophage with human EAC cell line SKGT. (A) Images of THP1 cells in control and polarised groups after co-cultured for 3 days without or with SKGT cells. (B) Quantification data from RT-Q-PCR show the expressions of cytokines/chemokines (n=3 individual experiments). (C) Representative images of EAC SKGT cells after migration and invasion. (D) Quantification data from migration and invasion experiments (n=5 individual experiments).
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fig4: Induction of THP1 monocyte into M2-like macrophage with human EAC cell line SKGT. (A) Images of THP1 cells in control and polarised groups after co-cultured for 3 days without or with SKGT cells. (B) Quantification data from RT-Q-PCR show the expressions of cytokines/chemokines (n=3 individual experiments). (C) Representative images of EAC SKGT cells after migration and invasion. (D) Quantification data from migration and invasion experiments (n=5 individual experiments).

Mentions: To explore the potential mechanisms underlying the correlation of M2 macrophage with nodal spread, in this study, we tested if EAC cells could polarise monocytes into M2-like macrophages, and then evaluated if polarised macrophages promoted migration and invasion of EAC cells. After co-culture of EAC cell line, SKGT cells, with THP1 monocytes for 3 days, macrophage morphology was clearly seen (Figure 4A). The expression profile of chemokines/cytokines was used to characterise subtype macrophage, which included CD11b, IL-10, IL-12, CCL17, CCL18, CD206, and CD163. Compared with untreated THP1 cells, expressions of IL-10, CD163, CD206, and CCL18 were increased in SKGT-induced THP1 cells (Figure 4B), suggesting polarised THP1 cells displayed M2-like phenotype. High expression of CCL18 indicated the macrophage subtype was close to recently reported M2d-like phenotype (Wang et al, 2010). Subsequently, we continued to employ the co-culture system to investigate if M2d-like macrophage induced by SKGT cells promoted EAC cell migration and invasion. The data showed that M2d-like macrophage not only stimulated EAC cell migration, but also increased cancer cell invasion (Figures 4C and D).


Macrophage subtype predicts lymph node metastasis in oesophageal adenocarcinoma and promotes cancer cell invasion in vitro
Induction of THP1 monocyte into M2-like macrophage with human EAC cell line SKGT. (A) Images of THP1 cells in control and polarised groups after co-cultured for 3 days without or with SKGT cells. (B) Quantification data from RT-Q-PCR show the expressions of cytokines/chemokines (n=3 individual experiments). (C) Representative images of EAC SKGT cells after migration and invasion. (D) Quantification data from migration and invasion experiments (n=5 individual experiments).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4559839&req=5

fig4: Induction of THP1 monocyte into M2-like macrophage with human EAC cell line SKGT. (A) Images of THP1 cells in control and polarised groups after co-cultured for 3 days without or with SKGT cells. (B) Quantification data from RT-Q-PCR show the expressions of cytokines/chemokines (n=3 individual experiments). (C) Representative images of EAC SKGT cells after migration and invasion. (D) Quantification data from migration and invasion experiments (n=5 individual experiments).
Mentions: To explore the potential mechanisms underlying the correlation of M2 macrophage with nodal spread, in this study, we tested if EAC cells could polarise monocytes into M2-like macrophages, and then evaluated if polarised macrophages promoted migration and invasion of EAC cells. After co-culture of EAC cell line, SKGT cells, with THP1 monocytes for 3 days, macrophage morphology was clearly seen (Figure 4A). The expression profile of chemokines/cytokines was used to characterise subtype macrophage, which included CD11b, IL-10, IL-12, CCL17, CCL18, CD206, and CD163. Compared with untreated THP1 cells, expressions of IL-10, CD163, CD206, and CCL18 were increased in SKGT-induced THP1 cells (Figure 4B), suggesting polarised THP1 cells displayed M2-like phenotype. High expression of CCL18 indicated the macrophage subtype was close to recently reported M2d-like phenotype (Wang et al, 2010). Subsequently, we continued to employ the co-culture system to investigate if M2d-like macrophage induced by SKGT cells promoted EAC cell migration and invasion. The data showed that M2d-like macrophage not only stimulated EAC cell migration, but also increased cancer cell invasion (Figures 4C and D).

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Currently, there is a lack of ideal biomarkers for predicting nodal status in preoperative stage of oesophageal adenocarcinoma (EAC) to aid optimising therapeutic options. We studied the potential of applying subtype macrophages to predict lymph node metastasis and prognosis in EAC.

Material and methods:: Fifty-three EAC resection specimens were immunostained with CD68, CD40 (M1), and CD163 (M2). Lymphatic vessel density (LVD) was estimated with the staining of D2-40. Subsequently, we tested if M2d macrophage could promote EAC cell migration and invasion.

Results:: In EAC without neoadjuvant treatment, an increase in M2-like macrophage was associated with poor patient survival, independent of the locations of macrophages in tumour. The M2/M1 ratio that represented the balance between M2- and M1-like macrophages was significantly higher in nodal-positive EACs than that in nodal-negative EACs, and inversely correlated with patient overall survival. The M2/M1 ratio was not related to LVD. EAC cell polarised THP1 cell into M2d-like macrophage, which promoted EAC cell migration and invasion. Neoadjuvant therapy appeared to diminish the correlation between the M2/M1 ratio and survival.

Conclusions:: The ratio of M2/M1 macrophage may serve as a sensitive marker to predict lymph node metastasis and poor prognosis in EAC without neoadjuvant therapy. M2d macrophage may have important roles in EAC metastasis.

No MeSH data available.


Related in: MedlinePlus