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Thrombospondin-1 (TSP-1) Stimulates Expression of Integrin alpha6 in Human Breast Carcinoma Cells: A Downstream Modulator of TSP-1-Induced Cellular Adhesion.

John AS, Rothman VL, Tuszynski GP - J Oncol (2010)

Bottom Line: This paper reports the novel finding that TSP-1 upregulates integrin alpha6 subunit in human keratinocytes and human breast cancer cells resulting in increased cell adhesion and tumor cell invasion.The effect of TSP-1 on alpha6 subunit expression was examined in human keratinocytes and breast adenocarcinoma cell lines (MDA-MB-231) treated with TSP-1 and in TSP-1 stably transfected breast cancer cells.These data suggest that TSP-1 plays an integral role in the attachment of cells to the ECM facilitating cell motility and angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Division of Pediatric Cardiology, Children's National Medical Center, George Washington University, Washington, DC 20052, USA.

ABSTRACT
Thrombospondin-1 (TSP-1) is involved in a variety of different cellular processes including cell adhesion, tumor progression, and angiogenesis. This paper reports the novel finding that TSP-1 upregulates integrin alpha6 subunit in human keratinocytes and human breast cancer cells resulting in increased cell adhesion and tumor cell invasion. The effect of TSP-1 on alpha6 subunit expression was examined in human keratinocytes and breast adenocarcinoma cell lines (MDA-MB-231) treated with TSP-1 and in TSP-1 stably transfected breast cancer cells. TSP-1 upregulated alpha6 message and protein in these cells as revealed by differential display, Northern and Western blot analysis and immunohistochemical localization studies. The increased expression of alpha6 was shown to mediate adhesion and invasion of these cells to laminin, a major component of the basement membrane and extracellular matrix (ECM). These data suggest that TSP-1 plays an integral role in the attachment of cells to the ECM facilitating cell motility and angiogenesis.

No MeSH data available.


Related in: MedlinePlus

Anti-TSP-1 antibody inhibition of integrin α6 production in TSP-1 stably transfected cells. Cells were grown in six well chamber slides in either serum-free media or media containing either 10 μg/mL control IgG or 10 μg/mL goat antihuman TSP-1 IgG, fixed, and stained with rat α6 integrin IgG as described in Section 2. Cells were photographed at 200X magnification. (a) TH5 cells (vector control).  (b) TH26 cells (high TSP-1 producer).  (c) TH26 cells plus anti-TSP-1 antibody.  (d) TH26 cells plus control IgG.
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Related In: Results  -  Collection


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fig5: Anti-TSP-1 antibody inhibition of integrin α6 production in TSP-1 stably transfected cells. Cells were grown in six well chamber slides in either serum-free media or media containing either 10 μg/mL control IgG or 10 μg/mL goat antihuman TSP-1 IgG, fixed, and stained with rat α6 integrin IgG as described in Section 2. Cells were photographed at 200X magnification. (a) TH5 cells (vector control). (b) TH26 cells (high TSP-1 producer). (c) TH26 cells plus anti-TSP-1 antibody. (d) TH26 cells plus control IgG.

Mentions: To determine if the staining observed in the stably transfected cells was specific to TSP-1, the TH26 cells were treated with either 10 μg/mL of polyclonal goat anti-TSP-1 IgG or 10 μg/mL of goat IgG and grown for an additional 24 hours. Immunohistochemical staining showed decreased expression of integrin α6 in the TH26 cells treated with the anti-TSP-I IgG comparable to the TH5 vector control, while the control antibody IgG-treated cells and untreated cells showed significant α6 staining (Figure 5). When the same experiment was repeated using an antitype 1 repeat TSP-1 antibody, there was no blocking effect (data not shown). These results suggest that endogenously produced TSP-1 specifically induces integrin α6 upregulation through domains other than the type 1 repeat domain of TSP-1.


Thrombospondin-1 (TSP-1) Stimulates Expression of Integrin alpha6 in Human Breast Carcinoma Cells: A Downstream Modulator of TSP-1-Induced Cellular Adhesion.

John AS, Rothman VL, Tuszynski GP - J Oncol (2010)

Anti-TSP-1 antibody inhibition of integrin α6 production in TSP-1 stably transfected cells. Cells were grown in six well chamber slides in either serum-free media or media containing either 10 μg/mL control IgG or 10 μg/mL goat antihuman TSP-1 IgG, fixed, and stained with rat α6 integrin IgG as described in Section 2. Cells were photographed at 200X magnification. (a) TH5 cells (vector control).  (b) TH26 cells (high TSP-1 producer).  (c) TH26 cells plus anti-TSP-1 antibody.  (d) TH26 cells plus control IgG.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2902750&req=5

fig5: Anti-TSP-1 antibody inhibition of integrin α6 production in TSP-1 stably transfected cells. Cells were grown in six well chamber slides in either serum-free media or media containing either 10 μg/mL control IgG or 10 μg/mL goat antihuman TSP-1 IgG, fixed, and stained with rat α6 integrin IgG as described in Section 2. Cells were photographed at 200X magnification. (a) TH5 cells (vector control). (b) TH26 cells (high TSP-1 producer). (c) TH26 cells plus anti-TSP-1 antibody. (d) TH26 cells plus control IgG.
Mentions: To determine if the staining observed in the stably transfected cells was specific to TSP-1, the TH26 cells were treated with either 10 μg/mL of polyclonal goat anti-TSP-1 IgG or 10 μg/mL of goat IgG and grown for an additional 24 hours. Immunohistochemical staining showed decreased expression of integrin α6 in the TH26 cells treated with the anti-TSP-I IgG comparable to the TH5 vector control, while the control antibody IgG-treated cells and untreated cells showed significant α6 staining (Figure 5). When the same experiment was repeated using an antitype 1 repeat TSP-1 antibody, there was no blocking effect (data not shown). These results suggest that endogenously produced TSP-1 specifically induces integrin α6 upregulation through domains other than the type 1 repeat domain of TSP-1.

Bottom Line: This paper reports the novel finding that TSP-1 upregulates integrin alpha6 subunit in human keratinocytes and human breast cancer cells resulting in increased cell adhesion and tumor cell invasion.The effect of TSP-1 on alpha6 subunit expression was examined in human keratinocytes and breast adenocarcinoma cell lines (MDA-MB-231) treated with TSP-1 and in TSP-1 stably transfected breast cancer cells.These data suggest that TSP-1 plays an integral role in the attachment of cells to the ECM facilitating cell motility and angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Division of Pediatric Cardiology, Children's National Medical Center, George Washington University, Washington, DC 20052, USA.

ABSTRACT
Thrombospondin-1 (TSP-1) is involved in a variety of different cellular processes including cell adhesion, tumor progression, and angiogenesis. This paper reports the novel finding that TSP-1 upregulates integrin alpha6 subunit in human keratinocytes and human breast cancer cells resulting in increased cell adhesion and tumor cell invasion. The effect of TSP-1 on alpha6 subunit expression was examined in human keratinocytes and breast adenocarcinoma cell lines (MDA-MB-231) treated with TSP-1 and in TSP-1 stably transfected breast cancer cells. TSP-1 upregulated alpha6 message and protein in these cells as revealed by differential display, Northern and Western blot analysis and immunohistochemical localization studies. The increased expression of alpha6 was shown to mediate adhesion and invasion of these cells to laminin, a major component of the basement membrane and extracellular matrix (ECM). These data suggest that TSP-1 plays an integral role in the attachment of cells to the ECM facilitating cell motility and angiogenesis.

No MeSH data available.


Related in: MedlinePlus