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Chemokine CXCL13 is overexpressed in the tumour tissue and in the peripheral blood of breast cancer patients.

Panse J, Friedrichs K, Marx A, Hildebrandt Y, Luetkens T, Barrels K, Horn C, Stahl T, Cao Y, Milde-Langosch K, Niendorf A, Kröger N, Wenzel S, Leuwer R, Bokemeyer C, Hegewisch-Becker S, Atanackovic D - Br. J. Cancer (2008)

Bottom Line: Levels of CXCR5, the receptor for CXCL13, were low in malignant and healthy breast tissues, and surface expression was not detected in vitro.Finally, we detected significantly elevated serum concentrations of CXCL13 in patients with metastatic disease (n = 54) as compared with controls (n = 44) and disease-free patients (n = 48).In conclusion, CXCL13 is overexpressed within breast cancer tissues, and increased serum levels of this cytokine can be found in breast cancer patients with metastatic disease pointing to a role of CXCL13 in the progression of breast cancer, suggesting that CXCL13 might serve as a useful therapeutic target and/or diagnostic marker in this malignancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology/Hematology, Center of Oncology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg 20246, Germany.

ABSTRACT
The abilities of chemokines in orchestrating cellular migration are utilised by different (patho-)biological networks including malignancies. However, except for CXCR4/CXCL12, little is known about the relation between tumour-related chemokine expression and the development and progression of solid tumours like breast cancer. In this study, microarray analyses revealed the overexpression of chemokine CXCL13 in breast cancer specimens. This finding was confirmed by real-time polymerase chain reaction in a larger set of samples (n = 34) and cell lines, and was validated on the protein level performing Western blot, ELISA, and immunohistochemistry. Levels of CXCR5, the receptor for CXCL13, were low in malignant and healthy breast tissues, and surface expression was not detected in vitro. However, we observed a strong (P = 0.0004) correlation between the expressions of CXCL13 and CXCR5 in breast cancer tissues, indicating a biologically relevant role of CXCR5 in vivo. Finally, we detected significantly elevated serum concentrations of CXCL13 in patients with metastatic disease (n = 54) as compared with controls (n = 44) and disease-free patients (n = 48). In conclusion, CXCL13 is overexpressed within breast cancer tissues, and increased serum levels of this cytokine can be found in breast cancer patients with metastatic disease pointing to a role of CXCL13 in the progression of breast cancer, suggesting that CXCL13 might serve as a useful therapeutic target and/or diagnostic marker in this malignancy.

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CXCL13 and its receptor CXCR5 are expressed intracellularly in breast cancer cell lines. (A) Expressions of chemokine CXCL13 and its receptor CXCR5 were examined in six breast cancer cell lines applying conventional RT–PCR (upper rows) and Western blot (lower rows). Housekeeping genes GAPDH and β-actin served as internal controls. (B) Breast cancer cell lines ZR-75, BT-10, and MCF-7 were examined regarding the protein expression of CXCR5 using flow cytometry. Histograms indicate staining intensity applying anti-CXCR5 antibody (black) or an appropriate isotype control (grey). (C) Seven breast cancer cell lines were cultured for 72 h in complete medium with or without activating cytokine TNF-α or INF-γ. mRNA levels of CXCL13 and CXCR5 were evaluated at baseline as well as after 24, 48, and 72 h applying real-time PCR. At the same time points, the concentration of CXCL13 protein in the culture supernatant was quantified using an ELISA. Cell surface expression of CXCR5 protein was evaluated by flow cytometry.
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fig3: CXCL13 and its receptor CXCR5 are expressed intracellularly in breast cancer cell lines. (A) Expressions of chemokine CXCL13 and its receptor CXCR5 were examined in six breast cancer cell lines applying conventional RT–PCR (upper rows) and Western blot (lower rows). Housekeeping genes GAPDH and β-actin served as internal controls. (B) Breast cancer cell lines ZR-75, BT-10, and MCF-7 were examined regarding the protein expression of CXCR5 using flow cytometry. Histograms indicate staining intensity applying anti-CXCR5 antibody (black) or an appropriate isotype control (grey). (C) Seven breast cancer cell lines were cultured for 72 h in complete medium with or without activating cytokine TNF-α or INF-γ. mRNA levels of CXCL13 and CXCR5 were evaluated at baseline as well as after 24, 48, and 72 h applying real-time PCR. At the same time points, the concentration of CXCL13 protein in the culture supernatant was quantified using an ELISA. Cell surface expression of CXCR5 protein was evaluated by flow cytometry.

Mentions: Given the significant overexpression of CXCL13 at the mRNA and protein levels in breast cancer tumour samples, we sought to determine the expression patterns of the chemokine ligand and its receptor in human breast cancer cell lines. We analysed six different breast cancer cell lines and found RNA and protein expressions of CXCL13 in all but one and CXCR5 expression in four of six cell lines (Figure 3A). Analysing the breast cancer lines by flow cytometry, we failed to detect a significant surface expression of the chemokine receptor, however, by performing intracellular staining, we observed strong expression of CXCR5 in the cytoplasma of cell lines ZR-75, BT-20, and MCF-7 (Figure 3B).


Chemokine CXCL13 is overexpressed in the tumour tissue and in the peripheral blood of breast cancer patients.

Panse J, Friedrichs K, Marx A, Hildebrandt Y, Luetkens T, Barrels K, Horn C, Stahl T, Cao Y, Milde-Langosch K, Niendorf A, Kröger N, Wenzel S, Leuwer R, Bokemeyer C, Hegewisch-Becker S, Atanackovic D - Br. J. Cancer (2008)

CXCL13 and its receptor CXCR5 are expressed intracellularly in breast cancer cell lines. (A) Expressions of chemokine CXCL13 and its receptor CXCR5 were examined in six breast cancer cell lines applying conventional RT–PCR (upper rows) and Western blot (lower rows). Housekeeping genes GAPDH and β-actin served as internal controls. (B) Breast cancer cell lines ZR-75, BT-10, and MCF-7 were examined regarding the protein expression of CXCR5 using flow cytometry. Histograms indicate staining intensity applying anti-CXCR5 antibody (black) or an appropriate isotype control (grey). (C) Seven breast cancer cell lines were cultured for 72 h in complete medium with or without activating cytokine TNF-α or INF-γ. mRNA levels of CXCL13 and CXCR5 were evaluated at baseline as well as after 24, 48, and 72 h applying real-time PCR. At the same time points, the concentration of CXCL13 protein in the culture supernatant was quantified using an ELISA. Cell surface expression of CXCR5 protein was evaluated by flow cytometry.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2538749&req=5

fig3: CXCL13 and its receptor CXCR5 are expressed intracellularly in breast cancer cell lines. (A) Expressions of chemokine CXCL13 and its receptor CXCR5 were examined in six breast cancer cell lines applying conventional RT–PCR (upper rows) and Western blot (lower rows). Housekeeping genes GAPDH and β-actin served as internal controls. (B) Breast cancer cell lines ZR-75, BT-10, and MCF-7 were examined regarding the protein expression of CXCR5 using flow cytometry. Histograms indicate staining intensity applying anti-CXCR5 antibody (black) or an appropriate isotype control (grey). (C) Seven breast cancer cell lines were cultured for 72 h in complete medium with or without activating cytokine TNF-α or INF-γ. mRNA levels of CXCL13 and CXCR5 were evaluated at baseline as well as after 24, 48, and 72 h applying real-time PCR. At the same time points, the concentration of CXCL13 protein in the culture supernatant was quantified using an ELISA. Cell surface expression of CXCR5 protein was evaluated by flow cytometry.
Mentions: Given the significant overexpression of CXCL13 at the mRNA and protein levels in breast cancer tumour samples, we sought to determine the expression patterns of the chemokine ligand and its receptor in human breast cancer cell lines. We analysed six different breast cancer cell lines and found RNA and protein expressions of CXCL13 in all but one and CXCR5 expression in four of six cell lines (Figure 3A). Analysing the breast cancer lines by flow cytometry, we failed to detect a significant surface expression of the chemokine receptor, however, by performing intracellular staining, we observed strong expression of CXCR5 in the cytoplasma of cell lines ZR-75, BT-20, and MCF-7 (Figure 3B).

Bottom Line: Levels of CXCR5, the receptor for CXCL13, were low in malignant and healthy breast tissues, and surface expression was not detected in vitro.Finally, we detected significantly elevated serum concentrations of CXCL13 in patients with metastatic disease (n = 54) as compared with controls (n = 44) and disease-free patients (n = 48).In conclusion, CXCL13 is overexpressed within breast cancer tissues, and increased serum levels of this cytokine can be found in breast cancer patients with metastatic disease pointing to a role of CXCL13 in the progression of breast cancer, suggesting that CXCL13 might serve as a useful therapeutic target and/or diagnostic marker in this malignancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology/Hematology, Center of Oncology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg 20246, Germany.

ABSTRACT
The abilities of chemokines in orchestrating cellular migration are utilised by different (patho-)biological networks including malignancies. However, except for CXCR4/CXCL12, little is known about the relation between tumour-related chemokine expression and the development and progression of solid tumours like breast cancer. In this study, microarray analyses revealed the overexpression of chemokine CXCL13 in breast cancer specimens. This finding was confirmed by real-time polymerase chain reaction in a larger set of samples (n = 34) and cell lines, and was validated on the protein level performing Western blot, ELISA, and immunohistochemistry. Levels of CXCR5, the receptor for CXCL13, were low in malignant and healthy breast tissues, and surface expression was not detected in vitro. However, we observed a strong (P = 0.0004) correlation between the expressions of CXCL13 and CXCR5 in breast cancer tissues, indicating a biologically relevant role of CXCR5 in vivo. Finally, we detected significantly elevated serum concentrations of CXCL13 in patients with metastatic disease (n = 54) as compared with controls (n = 44) and disease-free patients (n = 48). In conclusion, CXCL13 is overexpressed within breast cancer tissues, and increased serum levels of this cytokine can be found in breast cancer patients with metastatic disease pointing to a role of CXCL13 in the progression of breast cancer, suggesting that CXCL13 might serve as a useful therapeutic target and/or diagnostic marker in this malignancy.

Show MeSH
Related in: MedlinePlus