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The significance of the co-existence of osteopontin and tumor-associated macrophages in gastric cancer progression.

Lin CN, Wang CJ, Chao YJ, Lai MD, Shan YS - BMC Cancer (2015)

Bottom Line: M2 tumor-associated macrophages (M2-TAMs) also play an important role in cancer progression.In the co-culture study, OPN was able to attract U937 cells and skew them toward M2-TAMs through paracrine action.The M2-TAMs could increase the invasiveness of OPN+-AGS cells and the growth rate of xenograft of a mixture of co-cultured OPN+-AGS and U937 cells.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Osteopontin (OPN) can recruit macrophages to the site of inflammation and promote tumorigenesis. M2 tumor-associated macrophages (M2-TAMs) also play an important role in cancer progression. This study aimed to clarify the role of OPN and M2-TAMs co-existence in gastric cancer.

Methods: The levels of OPN and M2-TAMs were evaluated by immunohistochemical staining in 170 resected gastric cancer specimens that were collected from 1998 to 2012. M2-TAMs were identified by staining for an M2 marker, CD204. The prognostic significance and correlation between OPN and CD204 expression were analyzed. A co-culture system of OPN+-AGS and U937 cells was designed to study the effect of OPN on the skewing of macrophages toward M2-TAMs for gastric cancer progression in vitro and in vivo.

Results: Patients with high expression (>50%) of OPN or CD204 exhibited poor 5-year overall survival rates (48.61%, p = 0.0055, and 52.14%, p = 0.0498, respectively). A positive correlation was observed between OPN and CD204 expression and high co-expression of OPN and CD204 demonstrated poor 5-year overall survival rates (48.90%, p = 0.0131). In the co-culture study, OPN was able to attract U937 cells and skew them toward M2-TAMs through paracrine action. The M2-TAMs could increase the invasiveness of OPN+-AGS cells and the growth rate of xenograft of a mixture of co-cultured OPN+-AGS and U937 cells.

Conclusion: OPN can skew macrophages toward M2-TAMs during gastric cancer progression. The co-existence of OPN and infiltrating M2-TAMs correlates with disease progression and poor survival and thus can serve as a prognostic marker in gastric cancer.

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Related in: MedlinePlus

The co-expression of OPN and M2-TAMs promotes gastric tumorigenesis. (a) The expression of M1/M2 markers in U937 cells was analyzed after co-culture treatment by real-time PCR. LPS-treated U937 cells expressed IL-1 as M1 macrophages, and U937 cells alone were used as a negative control. TAMcli cells isolated from clinical specimens were used as a positive control. After co-culture with OPN+-AGS cells, the U937 cells differentiated into M2-TAMs expressing high levels of CD204 and IL-10. (b) After 72 hours of co-culture with U937 or TAMcli cells, the invasiveness of the OPN+-AGS cells increased but that of OPN-shRNA AGS cells did not. The increased cell invasiveness was also blocked by an OPN monoclonal antibody. (c) The mixture of co-cultured OPN+-AGS and U937 cells inoculated into nude mice showed poor survival compared with a mixture of co-cultured OPN-shRNA AGS and U937 cells or OPN+-AGS cells alone (p = 0.0498). (d) The neovascularization in the xenografts was shown by double-staining with anti-CD31 and anti-αSMA antibodies. Less neovascularization with normal vascular structure was found in xenografts of a mixture of co-cultured OPN-shRNA AGS and U937 cells.
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Fig5: The co-expression of OPN and M2-TAMs promotes gastric tumorigenesis. (a) The expression of M1/M2 markers in U937 cells was analyzed after co-culture treatment by real-time PCR. LPS-treated U937 cells expressed IL-1 as M1 macrophages, and U937 cells alone were used as a negative control. TAMcli cells isolated from clinical specimens were used as a positive control. After co-culture with OPN+-AGS cells, the U937 cells differentiated into M2-TAMs expressing high levels of CD204 and IL-10. (b) After 72 hours of co-culture with U937 or TAMcli cells, the invasiveness of the OPN+-AGS cells increased but that of OPN-shRNA AGS cells did not. The increased cell invasiveness was also blocked by an OPN monoclonal antibody. (c) The mixture of co-cultured OPN+-AGS and U937 cells inoculated into nude mice showed poor survival compared with a mixture of co-cultured OPN-shRNA AGS and U937 cells or OPN+-AGS cells alone (p = 0.0498). (d) The neovascularization in the xenografts was shown by double-staining with anti-CD31 and anti-αSMA antibodies. Less neovascularization with normal vascular structure was found in xenografts of a mixture of co-cultured OPN-shRNA AGS and U937 cells.

Mentions: After being co-cultured with OPN+-AGS cells, the U937 cell demonstrated significantly increased mRNA levels of CD204 and IL-10, as in TAMcli cells, when compared with cells co-cultured with OPN-shRNA AGS cells, stimulated with LPS (to become M1 macrophages exhibiting increased mRNA levels of IL-1) or treated with an OPN monoclonal antibody (Figure 5a). This result proved that OPN was able to recruit monocytes and skewed them toward a M2-TAMs phenotype in gastric cancer microenvironments.Figure 5


The significance of the co-existence of osteopontin and tumor-associated macrophages in gastric cancer progression.

Lin CN, Wang CJ, Chao YJ, Lai MD, Shan YS - BMC Cancer (2015)

The co-expression of OPN and M2-TAMs promotes gastric tumorigenesis. (a) The expression of M1/M2 markers in U937 cells was analyzed after co-culture treatment by real-time PCR. LPS-treated U937 cells expressed IL-1 as M1 macrophages, and U937 cells alone were used as a negative control. TAMcli cells isolated from clinical specimens were used as a positive control. After co-culture with OPN+-AGS cells, the U937 cells differentiated into M2-TAMs expressing high levels of CD204 and IL-10. (b) After 72 hours of co-culture with U937 or TAMcli cells, the invasiveness of the OPN+-AGS cells increased but that of OPN-shRNA AGS cells did not. The increased cell invasiveness was also blocked by an OPN monoclonal antibody. (c) The mixture of co-cultured OPN+-AGS and U937 cells inoculated into nude mice showed poor survival compared with a mixture of co-cultured OPN-shRNA AGS and U937 cells or OPN+-AGS cells alone (p = 0.0498). (d) The neovascularization in the xenografts was shown by double-staining with anti-CD31 and anti-αSMA antibodies. Less neovascularization with normal vascular structure was found in xenografts of a mixture of co-cultured OPN-shRNA AGS and U937 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4384326&req=5

Fig5: The co-expression of OPN and M2-TAMs promotes gastric tumorigenesis. (a) The expression of M1/M2 markers in U937 cells was analyzed after co-culture treatment by real-time PCR. LPS-treated U937 cells expressed IL-1 as M1 macrophages, and U937 cells alone were used as a negative control. TAMcli cells isolated from clinical specimens were used as a positive control. After co-culture with OPN+-AGS cells, the U937 cells differentiated into M2-TAMs expressing high levels of CD204 and IL-10. (b) After 72 hours of co-culture with U937 or TAMcli cells, the invasiveness of the OPN+-AGS cells increased but that of OPN-shRNA AGS cells did not. The increased cell invasiveness was also blocked by an OPN monoclonal antibody. (c) The mixture of co-cultured OPN+-AGS and U937 cells inoculated into nude mice showed poor survival compared with a mixture of co-cultured OPN-shRNA AGS and U937 cells or OPN+-AGS cells alone (p = 0.0498). (d) The neovascularization in the xenografts was shown by double-staining with anti-CD31 and anti-αSMA antibodies. Less neovascularization with normal vascular structure was found in xenografts of a mixture of co-cultured OPN-shRNA AGS and U937 cells.
Mentions: After being co-cultured with OPN+-AGS cells, the U937 cell demonstrated significantly increased mRNA levels of CD204 and IL-10, as in TAMcli cells, when compared with cells co-cultured with OPN-shRNA AGS cells, stimulated with LPS (to become M1 macrophages exhibiting increased mRNA levels of IL-1) or treated with an OPN monoclonal antibody (Figure 5a). This result proved that OPN was able to recruit monocytes and skewed them toward a M2-TAMs phenotype in gastric cancer microenvironments.Figure 5

Bottom Line: M2 tumor-associated macrophages (M2-TAMs) also play an important role in cancer progression.In the co-culture study, OPN was able to attract U937 cells and skew them toward M2-TAMs through paracrine action.The M2-TAMs could increase the invasiveness of OPN+-AGS cells and the growth rate of xenograft of a mixture of co-cultured OPN+-AGS and U937 cells.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Osteopontin (OPN) can recruit macrophages to the site of inflammation and promote tumorigenesis. M2 tumor-associated macrophages (M2-TAMs) also play an important role in cancer progression. This study aimed to clarify the role of OPN and M2-TAMs co-existence in gastric cancer.

Methods: The levels of OPN and M2-TAMs were evaluated by immunohistochemical staining in 170 resected gastric cancer specimens that were collected from 1998 to 2012. M2-TAMs were identified by staining for an M2 marker, CD204. The prognostic significance and correlation between OPN and CD204 expression were analyzed. A co-culture system of OPN+-AGS and U937 cells was designed to study the effect of OPN on the skewing of macrophages toward M2-TAMs for gastric cancer progression in vitro and in vivo.

Results: Patients with high expression (>50%) of OPN or CD204 exhibited poor 5-year overall survival rates (48.61%, p = 0.0055, and 52.14%, p = 0.0498, respectively). A positive correlation was observed between OPN and CD204 expression and high co-expression of OPN and CD204 demonstrated poor 5-year overall survival rates (48.90%, p = 0.0131). In the co-culture study, OPN was able to attract U937 cells and skew them toward M2-TAMs through paracrine action. The M2-TAMs could increase the invasiveness of OPN+-AGS cells and the growth rate of xenograft of a mixture of co-cultured OPN+-AGS and U937 cells.

Conclusion: OPN can skew macrophages toward M2-TAMs during gastric cancer progression. The co-existence of OPN and infiltrating M2-TAMs correlates with disease progression and poor survival and thus can serve as a prognostic marker in gastric cancer.

Show MeSH
Related in: MedlinePlus