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CD26 expression on T cell lines increases SDF-1-alpha-mediated invasion.

Havre PA, Abe M, Urasaki Y, Ohnuma K, Morimoto C, Dang NH - Br. J. Cancer (2009)

Bottom Line: This process was regulated in part by the PI-3K and MEK1 pathways, as indicated by increased phosphorylation of p44/42 MAP kinase and Akt in the presence of SDF-1-alpha and the effect of their respective inhibitors on MMP-9 secretion and in vitro invasion.In addition, CD26-associated enhancement of SDF-1-alpha-induced invasion was decreased when CD45 was inhibited.Our results indicate that the expression of CD26 in T cell lines leads to increased SDF-1-alpha-mediated invasion in an in vitro system and that this is controlled in part by the PI-3K and MEK1 pathways.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, University of Florida, Gainesville, FL 32610, USA.

ABSTRACT

Background: CD26 is a multifunctional membrane-bound glycoprotein that regulates tumour growth in addition to its other activities. Because disease aggressiveness is correlated with CD26 expression in several T-cell malignancies, we decided to investigate the invasiveness of cells expressing different levels of CD26.

Methods: To assess CD26 involvement in cell invasion, we performed in vitro invasion assays with human T cell lines expressing different levels of CD26. These included the parental CD26-positive T-lymphoblast cell line HSB-2 and clones infected with a retrovirus expressing siRNA vectors that either targeted CD26 or encoded a missense siRNA, and the parental CD26-negative T-leukaemia cell line Jurkat and clones expressing CD26. CD26 expression in these cell lines was evaluated by flow cytometry and western immunoblotting. CXCR4 expression, phosphorylation of signalling kinases, and MMP-9 secretion were also evaluated by western immunoblotting, whereas MMP-9 activity and the effect of kinase and CD45 inhibitors on activity were measured by zymography of conditioned media.

Results: The presence of CD26 enhanced stromal-cell-derived factor-1-alpha (SDF-1-alpha)-mediated invasion of T cell lines. This process was regulated in part by the PI-3K and MEK1 pathways, as indicated by increased phosphorylation of p44/42 MAP kinase and Akt in the presence of SDF-1-alpha and the effect of their respective inhibitors on MMP-9 secretion and in vitro invasion. In addition, CD26-associated enhancement of SDF-1-alpha-induced invasion was decreased when CD45 was inhibited.

Conclusions: Our results indicate that the expression of CD26 in T cell lines leads to increased SDF-1-alpha-mediated invasion in an in vitro system and that this is controlled in part by the PI-3K and MEK1 pathways. The data also suggest that CD26 enhancement of invasion may be mediated by CD45, however, more studies are required to confirm this involvement.

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CXCR4 level is not dependant on CD26 expression and is not affected by SDF-1-α. Expression of CXCR4 in whole-cell lysates. Equal amounts of protein were run in each lane of a 7.5% gel, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. (A) Lanes contained HSB-2 parental cells, H1-2 (parental cells with control siRNA), and three CD26-depleted clones: 2E5, 2F8, and 2G9. (B) Lanes contained CD26-negative Jurkat parental cells, Neo (parental cells with empty vector), and wt1, a CD26-overexpressing clone. When SDF-1-α was present (+), it was at a final concentration of 10 n.
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fig3: CXCR4 level is not dependant on CD26 expression and is not affected by SDF-1-α. Expression of CXCR4 in whole-cell lysates. Equal amounts of protein were run in each lane of a 7.5% gel, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. (A) Lanes contained HSB-2 parental cells, H1-2 (parental cells with control siRNA), and three CD26-depleted clones: 2E5, 2F8, and 2G9. (B) Lanes contained CD26-negative Jurkat parental cells, Neo (parental cells with empty vector), and wt1, a CD26-overexpressing clone. When SDF-1-α was present (+), it was at a final concentration of 10 n.

Mentions: Stromal-cell-derived factor-1-α presence has been shown to upregulate CXCR4 expression in the prostate cancer cell line PC-3 (Kukreja et al, 2005). We therefore considered the possibility that there might be a difference in the expression of CXCR4 among the parental cells and clones differing in CD26 levels. Furthermore, the possibility that CD26-expressing cells might upregulate CXCR4 in response to SDF-1-α, whereas CD26-negative cells might be resistant to change was also evaluated. Cells were maintained overnight in SFM containing 0.1% BSA, and SDF-1-α was added the following day at a final concentration of 10 n. Whole-cell lysates were prepared and run on gels, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. Our data showed that CXCR4 level was not affected by SDF-1-α (Figure 3). In addition, CXCR4 expression did not vary significantly among parental cells expressing CD26 (HSB-2 and H1-2) and CD26-depleted clones (2E5, 2F8, and 2G9) or among CD26-negative parental Jurkat cells (Jurkat and Neo) and a CD26-overexpressing clone (wt1).


CD26 expression on T cell lines increases SDF-1-alpha-mediated invasion.

Havre PA, Abe M, Urasaki Y, Ohnuma K, Morimoto C, Dang NH - Br. J. Cancer (2009)

CXCR4 level is not dependant on CD26 expression and is not affected by SDF-1-α. Expression of CXCR4 in whole-cell lysates. Equal amounts of protein were run in each lane of a 7.5% gel, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. (A) Lanes contained HSB-2 parental cells, H1-2 (parental cells with control siRNA), and three CD26-depleted clones: 2E5, 2F8, and 2G9. (B) Lanes contained CD26-negative Jurkat parental cells, Neo (parental cells with empty vector), and wt1, a CD26-overexpressing clone. When SDF-1-α was present (+), it was at a final concentration of 10 n.
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Related In: Results  -  Collection

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fig3: CXCR4 level is not dependant on CD26 expression and is not affected by SDF-1-α. Expression of CXCR4 in whole-cell lysates. Equal amounts of protein were run in each lane of a 7.5% gel, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. (A) Lanes contained HSB-2 parental cells, H1-2 (parental cells with control siRNA), and three CD26-depleted clones: 2E5, 2F8, and 2G9. (B) Lanes contained CD26-negative Jurkat parental cells, Neo (parental cells with empty vector), and wt1, a CD26-overexpressing clone. When SDF-1-α was present (+), it was at a final concentration of 10 n.
Mentions: Stromal-cell-derived factor-1-α presence has been shown to upregulate CXCR4 expression in the prostate cancer cell line PC-3 (Kukreja et al, 2005). We therefore considered the possibility that there might be a difference in the expression of CXCR4 among the parental cells and clones differing in CD26 levels. Furthermore, the possibility that CD26-expressing cells might upregulate CXCR4 in response to SDF-1-α, whereas CD26-negative cells might be resistant to change was also evaluated. Cells were maintained overnight in SFM containing 0.1% BSA, and SDF-1-α was added the following day at a final concentration of 10 n. Whole-cell lysates were prepared and run on gels, transferred to nitrocellulose, and probed with anti-CXCR4 antibody. Our data showed that CXCR4 level was not affected by SDF-1-α (Figure 3). In addition, CXCR4 expression did not vary significantly among parental cells expressing CD26 (HSB-2 and H1-2) and CD26-depleted clones (2E5, 2F8, and 2G9) or among CD26-negative parental Jurkat cells (Jurkat and Neo) and a CD26-overexpressing clone (wt1).

Bottom Line: This process was regulated in part by the PI-3K and MEK1 pathways, as indicated by increased phosphorylation of p44/42 MAP kinase and Akt in the presence of SDF-1-alpha and the effect of their respective inhibitors on MMP-9 secretion and in vitro invasion.In addition, CD26-associated enhancement of SDF-1-alpha-induced invasion was decreased when CD45 was inhibited.Our results indicate that the expression of CD26 in T cell lines leads to increased SDF-1-alpha-mediated invasion in an in vitro system and that this is controlled in part by the PI-3K and MEK1 pathways.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, University of Florida, Gainesville, FL 32610, USA.

ABSTRACT

Background: CD26 is a multifunctional membrane-bound glycoprotein that regulates tumour growth in addition to its other activities. Because disease aggressiveness is correlated with CD26 expression in several T-cell malignancies, we decided to investigate the invasiveness of cells expressing different levels of CD26.

Methods: To assess CD26 involvement in cell invasion, we performed in vitro invasion assays with human T cell lines expressing different levels of CD26. These included the parental CD26-positive T-lymphoblast cell line HSB-2 and clones infected with a retrovirus expressing siRNA vectors that either targeted CD26 or encoded a missense siRNA, and the parental CD26-negative T-leukaemia cell line Jurkat and clones expressing CD26. CD26 expression in these cell lines was evaluated by flow cytometry and western immunoblotting. CXCR4 expression, phosphorylation of signalling kinases, and MMP-9 secretion were also evaluated by western immunoblotting, whereas MMP-9 activity and the effect of kinase and CD45 inhibitors on activity were measured by zymography of conditioned media.

Results: The presence of CD26 enhanced stromal-cell-derived factor-1-alpha (SDF-1-alpha)-mediated invasion of T cell lines. This process was regulated in part by the PI-3K and MEK1 pathways, as indicated by increased phosphorylation of p44/42 MAP kinase and Akt in the presence of SDF-1-alpha and the effect of their respective inhibitors on MMP-9 secretion and in vitro invasion. In addition, CD26-associated enhancement of SDF-1-alpha-induced invasion was decreased when CD45 was inhibited.

Conclusions: Our results indicate that the expression of CD26 in T cell lines leads to increased SDF-1-alpha-mediated invasion in an in vitro system and that this is controlled in part by the PI-3K and MEK1 pathways. The data also suggest that CD26 enhancement of invasion may be mediated by CD45, however, more studies are required to confirm this involvement.

Show MeSH
Related in: MedlinePlus