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The chemokine CXCL16 and its receptor, CXCR6, as markers and promoters of inflammation-associated cancers.

Darash-Yahana M, Gillespie JW, Hewitt SM, Chen YY, Maeda S, Stein I, Singh SP, Bedolla RB, Peled A, Troyer DA, Pikarsky E, Karin M, Farber JM - PLoS ONE (2009)

Bottom Line: Screening for 37 chemokines in prostate cancer cell lines and xenografts revealed CXCL16, the ligand for the receptor CXCR6, as the most consistently expressed chemokine.Immunohistochemistry and/or immunofluorescence and confocal imaging of 121 human prostate specimens showed that CXCL16 and CXCR6 were co-expressed, both on prostate cancer cells and adjacent T cells.We studied expression of CXCL16 in an additional 461 specimens covering 12 tumor types, and found that CXCL16 was expressed in multiple human cancers associated with inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Immunology, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, MD, USA. meravd@hadassah.org.il

ABSTRACT
Clinical observations and mouse models have suggested that inflammation can be pro-tumorigenic. Since chemokines are critical in leukocyte trafficking, we hypothesized that chemokines play essential roles in inflammation-associated cancers. Screening for 37 chemokines in prostate cancer cell lines and xenografts revealed CXCL16, the ligand for the receptor CXCR6, as the most consistently expressed chemokine. Immunohistochemistry and/or immunofluorescence and confocal imaging of 121 human prostate specimens showed that CXCL16 and CXCR6 were co-expressed, both on prostate cancer cells and adjacent T cells. Expression levels of CXCL16 and CXCR6 on cancer cells correlated with poor prognostic features including high-stage and high-grade, and expression also correlated with post-inflammatory changes in the cancer stroma as revealed by loss of alpha-smooth muscle actin. Moreover, CXCL16 enhanced the growth of CXCR6-expressing cancer and primary CD4 T cells. We studied expression of CXCL16 in an additional 461 specimens covering 12 tumor types, and found that CXCL16 was expressed in multiple human cancers associated with inflammation. Our study is the first to describe the expression of CXCL16/CXCR6 on both cancer cells and adjacent T cells in humans, and to demonstrate correlations between CXCL16 and CXCR6 vs. poor both prognostic features and reactive changes in cancer stoma. Taken together, our data suggest that CXCL16 and CXCR6 may mark cancers arising in an inflammatory milieu and mediate pro-tumorigenic effects of inflammation through direct effects on cancer cell growth and by inducing the migration and proliferation of tumor-associated leukocytes.

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Related in: MedlinePlus

Roles for CXCL16 and CXCR6 in prostate cancer stage, grade and cell growth.(A) Arrays containing prostate cancer tissue from 40 patients were stained for CXCL16 and CXCR6. Samples were scored for CXCL16 and CXCR6 expression as described in Materials and Methods. Samples were grouped according to stage or Gleason score (X-axes) as determined by the supplier, and mean scores for CXCL16 and CXCR6 expression were calculated. The number of samples (n) in each stage or Gleason score group is shown. Error bars show SEM. Cross-bars indicate comparisons that are significant. *, p<0.05 and ***, p<0.001. (B) PC3 cells were transfected with the CXCR6-YFP plasmid, cultured for 48 hours without chemokine (control, ctrl) or with 0.05–5 µg/ml CXCL16 and numbers of CXCR6-YFP+ (R6YFP+) vs. CXCR6YFP− (R6YFP−) cells were counted as described in Materials and Methods. Values were normalized to the control, untreated sample to yield the fold difference in cell number. Bars show means±SEM combined from three experiments. **, p<0.01 and ***, p<0.001 vs. control values. (C) Following transfection with the CXCR6-YFP plasmid, PC3 cells were cultured as in (B), without or with 5 µg/ml CXCL16. Left panel, gating is shown for separating untreated and CXCL16-treated cells into “low” (L) or “high” (H) for CXCR6-YFP. Percentages of cells in the high gates are shown. One representative experiment is shown out of three performed. Right panel, CXCR6-YFP+low (CXCR6+low) and CXCR6-YFP+high (CXCR6+high) cells from CXCL16-treated and control cultures were counted after 48 hours as in (B). Data were combined from three experiments. *, p<0.05 and ***, p<0.001 vs. untreated cells. (D) Confocal imaging displays anti-CXCL16 or anti-CXCR6 (488 tyramide) as green, anti-Ki67 (594 tyramide) as red, and nuclear staining with Hoechst 33258 as blue. Panel is representative of more than 15 cases.
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pone-0006695-g002: Roles for CXCL16 and CXCR6 in prostate cancer stage, grade and cell growth.(A) Arrays containing prostate cancer tissue from 40 patients were stained for CXCL16 and CXCR6. Samples were scored for CXCL16 and CXCR6 expression as described in Materials and Methods. Samples were grouped according to stage or Gleason score (X-axes) as determined by the supplier, and mean scores for CXCL16 and CXCR6 expression were calculated. The number of samples (n) in each stage or Gleason score group is shown. Error bars show SEM. Cross-bars indicate comparisons that are significant. *, p<0.05 and ***, p<0.001. (B) PC3 cells were transfected with the CXCR6-YFP plasmid, cultured for 48 hours without chemokine (control, ctrl) or with 0.05–5 µg/ml CXCL16 and numbers of CXCR6-YFP+ (R6YFP+) vs. CXCR6YFP− (R6YFP−) cells were counted as described in Materials and Methods. Values were normalized to the control, untreated sample to yield the fold difference in cell number. Bars show means±SEM combined from three experiments. **, p<0.01 and ***, p<0.001 vs. control values. (C) Following transfection with the CXCR6-YFP plasmid, PC3 cells were cultured as in (B), without or with 5 µg/ml CXCL16. Left panel, gating is shown for separating untreated and CXCL16-treated cells into “low” (L) or “high” (H) for CXCR6-YFP. Percentages of cells in the high gates are shown. One representative experiment is shown out of three performed. Right panel, CXCR6-YFP+low (CXCR6+low) and CXCR6-YFP+high (CXCR6+high) cells from CXCL16-treated and control cultures were counted after 48 hours as in (B). Data were combined from three experiments. *, p<0.05 and ***, p<0.001 vs. untreated cells. (D) Confocal imaging displays anti-CXCL16 or anti-CXCR6 (488 tyramide) as green, anti-Ki67 (594 tyramide) as red, and nuclear staining with Hoechst 33258 as blue. Panel is representative of more than 15 cases.

Mentions: The stage (I-IV) and grade (Gleason score) of prostate cancer are determined at diagnosis, and were reported by the supplier for the cases used in the arrays. Higher stage corresponds to greater anatomic extension of the cancer, and Gleason score describes histological characteristics that correlate with clinical outcome. Gleason scores of 2–4 are low-grade, scores 5–7 are moderate-grade, and scores 8–10 are high-grade prostate cancer. Analysis of the 40-case array for levels of staining for CXCL16 and CXCR6 revealed that their expression by the cancer cells correlated with both high-stage and high-grade prostate cancer (Figure 2A).


The chemokine CXCL16 and its receptor, CXCR6, as markers and promoters of inflammation-associated cancers.

Darash-Yahana M, Gillespie JW, Hewitt SM, Chen YY, Maeda S, Stein I, Singh SP, Bedolla RB, Peled A, Troyer DA, Pikarsky E, Karin M, Farber JM - PLoS ONE (2009)

Roles for CXCL16 and CXCR6 in prostate cancer stage, grade and cell growth.(A) Arrays containing prostate cancer tissue from 40 patients were stained for CXCL16 and CXCR6. Samples were scored for CXCL16 and CXCR6 expression as described in Materials and Methods. Samples were grouped according to stage or Gleason score (X-axes) as determined by the supplier, and mean scores for CXCL16 and CXCR6 expression were calculated. The number of samples (n) in each stage or Gleason score group is shown. Error bars show SEM. Cross-bars indicate comparisons that are significant. *, p<0.05 and ***, p<0.001. (B) PC3 cells were transfected with the CXCR6-YFP plasmid, cultured for 48 hours without chemokine (control, ctrl) or with 0.05–5 µg/ml CXCL16 and numbers of CXCR6-YFP+ (R6YFP+) vs. CXCR6YFP− (R6YFP−) cells were counted as described in Materials and Methods. Values were normalized to the control, untreated sample to yield the fold difference in cell number. Bars show means±SEM combined from three experiments. **, p<0.01 and ***, p<0.001 vs. control values. (C) Following transfection with the CXCR6-YFP plasmid, PC3 cells were cultured as in (B), without or with 5 µg/ml CXCL16. Left panel, gating is shown for separating untreated and CXCL16-treated cells into “low” (L) or “high” (H) for CXCR6-YFP. Percentages of cells in the high gates are shown. One representative experiment is shown out of three performed. Right panel, CXCR6-YFP+low (CXCR6+low) and CXCR6-YFP+high (CXCR6+high) cells from CXCL16-treated and control cultures were counted after 48 hours as in (B). Data were combined from three experiments. *, p<0.05 and ***, p<0.001 vs. untreated cells. (D) Confocal imaging displays anti-CXCL16 or anti-CXCR6 (488 tyramide) as green, anti-Ki67 (594 tyramide) as red, and nuclear staining with Hoechst 33258 as blue. Panel is representative of more than 15 cases.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2723911&req=5

pone-0006695-g002: Roles for CXCL16 and CXCR6 in prostate cancer stage, grade and cell growth.(A) Arrays containing prostate cancer tissue from 40 patients were stained for CXCL16 and CXCR6. Samples were scored for CXCL16 and CXCR6 expression as described in Materials and Methods. Samples were grouped according to stage or Gleason score (X-axes) as determined by the supplier, and mean scores for CXCL16 and CXCR6 expression were calculated. The number of samples (n) in each stage or Gleason score group is shown. Error bars show SEM. Cross-bars indicate comparisons that are significant. *, p<0.05 and ***, p<0.001. (B) PC3 cells were transfected with the CXCR6-YFP plasmid, cultured for 48 hours without chemokine (control, ctrl) or with 0.05–5 µg/ml CXCL16 and numbers of CXCR6-YFP+ (R6YFP+) vs. CXCR6YFP− (R6YFP−) cells were counted as described in Materials and Methods. Values were normalized to the control, untreated sample to yield the fold difference in cell number. Bars show means±SEM combined from three experiments. **, p<0.01 and ***, p<0.001 vs. control values. (C) Following transfection with the CXCR6-YFP plasmid, PC3 cells were cultured as in (B), without or with 5 µg/ml CXCL16. Left panel, gating is shown for separating untreated and CXCL16-treated cells into “low” (L) or “high” (H) for CXCR6-YFP. Percentages of cells in the high gates are shown. One representative experiment is shown out of three performed. Right panel, CXCR6-YFP+low (CXCR6+low) and CXCR6-YFP+high (CXCR6+high) cells from CXCL16-treated and control cultures were counted after 48 hours as in (B). Data were combined from three experiments. *, p<0.05 and ***, p<0.001 vs. untreated cells. (D) Confocal imaging displays anti-CXCL16 or anti-CXCR6 (488 tyramide) as green, anti-Ki67 (594 tyramide) as red, and nuclear staining with Hoechst 33258 as blue. Panel is representative of more than 15 cases.
Mentions: The stage (I-IV) and grade (Gleason score) of prostate cancer are determined at diagnosis, and were reported by the supplier for the cases used in the arrays. Higher stage corresponds to greater anatomic extension of the cancer, and Gleason score describes histological characteristics that correlate with clinical outcome. Gleason scores of 2–4 are low-grade, scores 5–7 are moderate-grade, and scores 8–10 are high-grade prostate cancer. Analysis of the 40-case array for levels of staining for CXCL16 and CXCR6 revealed that their expression by the cancer cells correlated with both high-stage and high-grade prostate cancer (Figure 2A).

Bottom Line: Screening for 37 chemokines in prostate cancer cell lines and xenografts revealed CXCL16, the ligand for the receptor CXCR6, as the most consistently expressed chemokine.Immunohistochemistry and/or immunofluorescence and confocal imaging of 121 human prostate specimens showed that CXCL16 and CXCR6 were co-expressed, both on prostate cancer cells and adjacent T cells.We studied expression of CXCL16 in an additional 461 specimens covering 12 tumor types, and found that CXCL16 was expressed in multiple human cancers associated with inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Immunology, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, MD, USA. meravd@hadassah.org.il

ABSTRACT
Clinical observations and mouse models have suggested that inflammation can be pro-tumorigenic. Since chemokines are critical in leukocyte trafficking, we hypothesized that chemokines play essential roles in inflammation-associated cancers. Screening for 37 chemokines in prostate cancer cell lines and xenografts revealed CXCL16, the ligand for the receptor CXCR6, as the most consistently expressed chemokine. Immunohistochemistry and/or immunofluorescence and confocal imaging of 121 human prostate specimens showed that CXCL16 and CXCR6 were co-expressed, both on prostate cancer cells and adjacent T cells. Expression levels of CXCL16 and CXCR6 on cancer cells correlated with poor prognostic features including high-stage and high-grade, and expression also correlated with post-inflammatory changes in the cancer stroma as revealed by loss of alpha-smooth muscle actin. Moreover, CXCL16 enhanced the growth of CXCR6-expressing cancer and primary CD4 T cells. We studied expression of CXCL16 in an additional 461 specimens covering 12 tumor types, and found that CXCL16 was expressed in multiple human cancers associated with inflammation. Our study is the first to describe the expression of CXCL16/CXCR6 on both cancer cells and adjacent T cells in humans, and to demonstrate correlations between CXCL16 and CXCR6 vs. poor both prognostic features and reactive changes in cancer stoma. Taken together, our data suggest that CXCL16 and CXCR6 may mark cancers arising in an inflammatory milieu and mediate pro-tumorigenic effects of inflammation through direct effects on cancer cell growth and by inducing the migration and proliferation of tumor-associated leukocytes.

Show MeSH
Related in: MedlinePlus