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Cell-line-specific stimulation of tumor cell aggressiveness by wound healing factors - a central role for STAT3.

Ekblad L, Lindgren G, Persson E, Kjellén E, Wennerberg J - BMC Cancer (2013)

Bottom Line: These effects were accompanied by a sharp increase in activation of signal transducer and activator of transcription 3 (STAT3).The involvement of c-Met or epidermal growth factor receptor family members was excluded, while the interleukin-6 receptor was found to be partly responsible for the activation of STAT3.In conclusion, we found cell-line-specific effects of wound healing factors on HNSCC, setting the stage for therapy development and predictive opportunities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, Lund University, Lund, Sweden. Lars.Ekblad@med.lu.se

ABSTRACT

Background: Local recurrence is a major factor affecting survival after treatment for head and neck squamous cell carcinoma (HNSCC). It is possible that the normal processes involved in wound healing after surgical removal of a primary tumor can boost the regrowth of residual cancer cells, thereby contributing to the recurrent growth. In this work, we collected human wound fluids and used them to investigate the effect of wound healing factors on HNSCC cell lines in vitro.

Methods: Wound fluids were collected from thyroidectomized patients diagnosed with benign disease and were included in assays of cell proliferation, migration, cell scattering, and invasion. The involvement of intracellular signaling pathways and membrane receptors were investigated by western blotting and the inclusion of specific inhibitors.

Results: One out of four cell lines was greatly stimulated in proliferation, migration, cell scattering, and invasion by the addition of wound fluid as compared with addition of fetal bovine or human serum. These effects were accompanied by a sharp increase in activation of signal transducer and activator of transcription 3 (STAT3). Inhibition of STAT3 activation abolished the wound fluid response, showing that STAT3 plays an important role in the wound healing response. Several of the observed phenotypic changes were epithelial-to-mesenchymal transition (EMT)-like, but the appropriate changes were not seen in any of the EMT markers investigated. The involvement of c-Met or epidermal growth factor receptor family members was excluded, while the interleukin-6 receptor was found to be partly responsible for the activation of STAT3.

Conclusions: In conclusion, we found cell-line-specific effects of wound healing factors on HNSCC, setting the stage for therapy development and predictive opportunities.

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Related in: MedlinePlus

The effect of HWF on invasive growth in Matrigel matrix. The cells were seeded in 90% Matrigel and 10% complete medium and left to form colonies. On day 0 the medium on top of the Matrigel matrix was changed to DMEM supplemented with 10% FBS, HS, or HWF. New medium was added every third day. The cells were photographed with a 10× objective.
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Figure 3: The effect of HWF on invasive growth in Matrigel matrix. The cells were seeded in 90% Matrigel and 10% complete medium and left to form colonies. On day 0 the medium on top of the Matrigel matrix was changed to DMEM supplemented with 10% FBS, HS, or HWF. New medium was added every third day. The cells were photographed with a 10× objective.

Mentions: HWF has a complex composition which probably includes chemotactic migration stimulating factors in addition to any possible invasion activators; this makes it difficult to design and interpret classical invasion assays with Matrigel-covered trans-well membranes. Instead, we chose to study the growth of the cell lines in three-dimensional Matrigel cultures. All four cell lines grew in spherical colonies when supplemented with 10% FBS in this matrix. Exchanging FBS for HWF drastically changed the growth of HN-7 cells; within the Matrigel matrix, these formed thread-like protrusions of cells contacting nearby colonies. After 1–2 weeks, these protrusions had formed a dynamic network of cells connecting the colonies throughout the matrix. Similar but much less pronounced effects were seen when HS was used instead of HWF (Figure 3).


Cell-line-specific stimulation of tumor cell aggressiveness by wound healing factors - a central role for STAT3.

Ekblad L, Lindgren G, Persson E, Kjellén E, Wennerberg J - BMC Cancer (2013)

The effect of HWF on invasive growth in Matrigel matrix. The cells were seeded in 90% Matrigel and 10% complete medium and left to form colonies. On day 0 the medium on top of the Matrigel matrix was changed to DMEM supplemented with 10% FBS, HS, or HWF. New medium was added every third day. The cells were photographed with a 10× objective.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3585883&req=5

Figure 3: The effect of HWF on invasive growth in Matrigel matrix. The cells were seeded in 90% Matrigel and 10% complete medium and left to form colonies. On day 0 the medium on top of the Matrigel matrix was changed to DMEM supplemented with 10% FBS, HS, or HWF. New medium was added every third day. The cells were photographed with a 10× objective.
Mentions: HWF has a complex composition which probably includes chemotactic migration stimulating factors in addition to any possible invasion activators; this makes it difficult to design and interpret classical invasion assays with Matrigel-covered trans-well membranes. Instead, we chose to study the growth of the cell lines in three-dimensional Matrigel cultures. All four cell lines grew in spherical colonies when supplemented with 10% FBS in this matrix. Exchanging FBS for HWF drastically changed the growth of HN-7 cells; within the Matrigel matrix, these formed thread-like protrusions of cells contacting nearby colonies. After 1–2 weeks, these protrusions had formed a dynamic network of cells connecting the colonies throughout the matrix. Similar but much less pronounced effects were seen when HS was used instead of HWF (Figure 3).

Bottom Line: These effects were accompanied by a sharp increase in activation of signal transducer and activator of transcription 3 (STAT3).The involvement of c-Met or epidermal growth factor receptor family members was excluded, while the interleukin-6 receptor was found to be partly responsible for the activation of STAT3.In conclusion, we found cell-line-specific effects of wound healing factors on HNSCC, setting the stage for therapy development and predictive opportunities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, Lund University, Lund, Sweden. Lars.Ekblad@med.lu.se

ABSTRACT

Background: Local recurrence is a major factor affecting survival after treatment for head and neck squamous cell carcinoma (HNSCC). It is possible that the normal processes involved in wound healing after surgical removal of a primary tumor can boost the regrowth of residual cancer cells, thereby contributing to the recurrent growth. In this work, we collected human wound fluids and used them to investigate the effect of wound healing factors on HNSCC cell lines in vitro.

Methods: Wound fluids were collected from thyroidectomized patients diagnosed with benign disease and were included in assays of cell proliferation, migration, cell scattering, and invasion. The involvement of intracellular signaling pathways and membrane receptors were investigated by western blotting and the inclusion of specific inhibitors.

Results: One out of four cell lines was greatly stimulated in proliferation, migration, cell scattering, and invasion by the addition of wound fluid as compared with addition of fetal bovine or human serum. These effects were accompanied by a sharp increase in activation of signal transducer and activator of transcription 3 (STAT3). Inhibition of STAT3 activation abolished the wound fluid response, showing that STAT3 plays an important role in the wound healing response. Several of the observed phenotypic changes were epithelial-to-mesenchymal transition (EMT)-like, but the appropriate changes were not seen in any of the EMT markers investigated. The involvement of c-Met or epidermal growth factor receptor family members was excluded, while the interleukin-6 receptor was found to be partly responsible for the activation of STAT3.

Conclusions: In conclusion, we found cell-line-specific effects of wound healing factors on HNSCC, setting the stage for therapy development and predictive opportunities.

Show MeSH
Related in: MedlinePlus