uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Interactions between Malignant Keratinocytes and Fibroblasts: Studies in Head and Neck Squamous Cell Carcinoma
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Plastic Surgery.
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Carcinoma growth requires a supportive tumor stroma. The concept of reciprocal interactions between tumor and stromal cells has become widely acknowledged and the connective tissue activation seen in the malignant process has been likened to that of a healing wound. Little is, however, known about the specific characteristics of these interactions, distinguishing them from the interplay occurring between epithelial and stromal cells in wound healing. In order to study differences in the humoral effects of malignant and benign epithelial cells on fibroblasts, we used an in vitro coculture model with human oral squamous cell carcinoma cells (SCC) or normal oral keratinocytes (NOK) on one side of a semi-permeable membrane and fibroblasts seeded in gels on the other. Pro-collagens α1(I) and α1(III) were more downregulated in NOK cocultures compared to SCC cocultures. IL-1α was identified as a major keratinocyte-derived soluble factor behind the effects observed. We concluded that SCC are less antifibrotic compared to NOK. There was also a differential expression among enzymes involved in ECM turnover. The urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) were both upregulated by NOK, but not by SCC. Here, rIL-1ra caused further upregulation of PAI-1. Global gene expression in fibroblasts was assessed using Affymetrix™ arrays. In total, 82 transcripts were considered differentially expressed; 52 were up- and 30 were downregulated in SCC compared to NOK cocultures. Among the differentially expressed genes there was an enrichment of genes related to collagens and to a nonspecific, innate-type response. The innate response marker pentraxin (PTX3) was upregulated by keratinocyte-derrived IL-1α in both NOK and SCC cocultures. We observed a considerably higher IL-1α / IL-1ra quotient in SCC cocultures, however, while PTX3 mRNA upregulation was higher in SCC cocultures, there was no difference in the level of PTX3 secreted protein. Taken together, we concluded that NOK and SCC regulate genes important for ECM composition and for the innate immune-response differentially. IL-1α was identified as one important mediator of the observed effects. In general, SCC appeared to be more profibrotic in their effects on fibroblasts. 

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. , 48 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 990
Keyword [en]
coculture, extracellular matrix, interleukin-1 alpha, tumor stroma, differential gene regulation, innate response
National Category
Surgery
Research subject
Plastic Surgery
Identifiers
URN: urn:nbn:se:uu:diva-221109ISBN: 978-91-554-8926-7 (print)OAI: oai:DiVA.org:uu-221109DiVA: diva2:709085
Public defence
2014-05-21, Skoogsalen, Akademiska sjukhuset, Uppsala, 09:15 (Swedish)
Opponent
Supervisors
Available from: 2014-04-29 Created: 2014-03-25 Last updated: 2014-06-30
List of papers
1. Interleukin-1-mediated effects of normal oral keratinocytes and head and neck squamous carcinoma cells on extracellular matrix related gene expression in fibroblasts
Open this publication in new window or tab >>Interleukin-1-mediated effects of normal oral keratinocytes and head and neck squamous carcinoma cells on extracellular matrix related gene expression in fibroblasts
Show others...
2012 (English)In: Oral Oncology, ISSN 1368-8375, E-ISSN 1879-0593, Vol. 48, no 12, 1236-1241 p.Article in journal (Refereed) Published
Abstract [en]

Objectives: The composition of tumor stroma and the activity of tumor associated fibroblasts are important for tumor growth. Interactions between carcinoma cells and fibroblasts regulate the turnover of extracellular matrix (ECM). Here, the in vitro effects of oral squamous cell carcinoma (SCC) cells (UT-SCC-30 and UT-SCC-87) on fibroblast expression of genes for ECM components and connective tissue growth factor (CTGF/CCN2), were compared to those of normal oral keratinocytes (NOK). Materials and Methods: Cocultures with fibroblasts in collagen gels and keratinocytes with the two cell types separated by a semi permeable membrane were used, and relative gene expression was measured with real-time PCR. Results: All investigated genes were regulated by NOK and the SCCs. The downregulation of pro-collagens alpha 1(I) and alpha 1(III) was more pronounced in cocultures with NOK, while the expression of CCN2 and fibronectin was downregulated by both NOK and the SCCs to a similar extent. UT-SCC-87, but not UT-SCC-30, secreted significantly more IL-1 alpha than NOK. A recombinant interleukin-1 receptor antagonist reversed many of the observed effects on fibroblast gene expression suggesting involvement of IL-1 in cocultures with NOK as well as with SCCs. Conclusion: The observed differential effects on fibroblast gene expression suggest that NOK are more antifibrotic compared to UT-SCC-30 and UT-SCC-87. These findings may contribute to a better understanding of the mechanisms behind ECM turnover in tumors.

Keyword
Cocultures, Extracellular matrix, Interleukin-1 alpha, Tumor stroma
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-188413 (URN)10.1016/j.oraloncology.2012.06.013 (DOI)000311151200007 ()
Available from: 2012-12-17 Created: 2012-12-17 Last updated: 2017-12-06Bibliographically approved
2. Keratinocytes and Head and Neck Squamous Cell Carcinoma Cells Regulate Urokinase-type Plasminogen Activator and Plasminogen Activator Inhibitor-1 in Fibroblasts
Open this publication in new window or tab >>Keratinocytes and Head and Neck Squamous Cell Carcinoma Cells Regulate Urokinase-type Plasminogen Activator and Plasminogen Activator Inhibitor-1 in Fibroblasts
Show others...
2013 (English)In: Anticancer Research, ISSN 0250-7005, E-ISSN 1791-7530, Vol. 33, no 8, 3113-3118 p.Article in journal (Refereed) Published
Abstract [en]

Background: To investigate possible differences in the effects of soluble factors from oral squamous cell carcinoma (SCC) cells (UT-SCC-87) and normal oral keratinocytes (NOK) on fibroblast expression of genes involved in tumor stroma turnover. Materials and Methods: Transwell co-cultures with fibroblasts in collagen gels, and SCC cells or NOK in inserts were carried out. Fibroblast gene expression was measured with real-time polymerase chain reaction (PCR). Results: The expression of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) was up-regulated in co-cultures with SCC cells but not with NOK. In contrast, both SCC cells and NOK regulated matrix metalloproteinase-1 (MMP1) and -3, and tissue inhibitor of metalloproteinases-2 (TIMP2) and -3 to a similar extent, while MMP2 and TIMP1 were largely unaffected. Interleukin 1 alpha (IL1 alpha) up-regulated both MMP1 and MMP3 and down-regulated PAI-1, TIMP2 and -3. Conclusion: SCC and NOK regulate fibroblast expression of genes involved in tumor stroma turnover differentially in vitro. These observations may contribute to a better understanding of the mechanisms behind extracellular matrix turnover in tumors.

Keyword
Head and neck cancer, keratinocytes, differential regulation, PAI-1, uPA
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-206561 (URN)000322559300018 ()
Available from: 2013-09-02 Created: 2013-09-02 Last updated: 2017-12-06Bibliographically approved
3. Differential gene regulation in fibroblasts in coculture with normal oral keratinocytes and head and neck squamous cell carcinoma cells
Open this publication in new window or tab >>Differential gene regulation in fibroblasts in coculture with normal oral keratinocytes and head and neck squamous cell carcinoma cells
Show others...
(English)Manuscript (preprint) (Other academic)
Keyword
microarray, global gene expression pattern, innate response
National Category
Surgery
Research subject
Plastic Surgery
Identifiers
urn:nbn:se:uu:diva-221115 (URN)
Available from: 2014-03-26 Created: 2014-03-25 Last updated: 2015-03-17
4. Normal oral keratinocytes and head and neck squamous carcinoma cells induce an innate response in fibroblasts
Open this publication in new window or tab >>Normal oral keratinocytes and head and neck squamous carcinoma cells induce an innate response in fibroblasts
Show others...
2016 (English)In: Anticancer Research, ISSN 0250-7005, E-ISSN 1791-7530, Vol. 36, no 5, 2131-2137 p.Article in journal (Other academic) Published
Abstract [en]

Background: Tumor stroma is similar to the connective tissue of chronic inflammation. The extracellular matrix of tumors is formed by cancer-associated fibroblasts that also modulate the inflammatory response. Materials and Methods: We studied the ability of oral keratinocytes (NOK) and oral squamous cell carcinoma cells (SCC) to induce an innate immune response in fibroblasts. Co-cultures with fibroblasts in collagen gels and keratinocytes in inserts were used. Pentraxin 3 (PTX3) was used as an indicator of an innate immune response. Results: SCC and NOK up-regulated fibroblast mRNA expression and protein release of PTX3. mRNA levels were more pronounced in cultures with malignant cells. The induction of PTX3 was abrogated by an interleukin-1 receptor antagonist Conclusion: Keratinocytes have the capacity to induce an interleukin-1-dependent innate immune response by fibroblasts in vitro. This could be important for subsequent fibroblast modulation of the inflammatory reaction in non-malignant and malignant disease processes.

Keyword
co-culture, pentraxin-3, extracellular matrix, interleukin-1 alpha, tumor stroma
National Category
Surgery Cancer and Oncology
Research subject
Plastic Surgery
Identifiers
urn:nbn:se:uu:diva-221123 (URN)000375456200010 ()27127114 (PubMedID)
Available from: 2014-03-26 Created: 2014-03-25 Last updated: 2017-12-05Bibliographically approved

Open Access in DiVA

fulltext(2401 kB)468 downloads
File information
File name FULLTEXT01.pdfFile size 2401 kBChecksum SHA-512
b6cf9b73ac9ab2e183822ea7a7f720ccd4274c7b460690109a6bbc4377fcdc7d90fc3d607c2b2d39178f351df74a1f66b8f59b22e6b9d9887431e39e4298b55e
Type fulltextMimetype application/pdf
Buy this publication >>

By organisation
Plastic Surgery
Surgery

Search outside of DiVA

GoogleGoogle Scholar
Total: 468 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 954 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf