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  • 1. Hellquist, Henrik B.
    et al.
    Sundelin, Kaarina
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    DiBacco, A.
    Department of Pathology and Laboratory Medicine, European Institute of Oncology, Milan University School of Medicine, Milan, Italy.
    Tytor, Maria
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences.
    Manzotti, Michela
    Department of Pathology and Laboratory Medicine, European Institute of Oncology, Milan University School of Medicine, Milan, Italy.
    Viale, Giuseppe J.
    Department of Pathology and Laboratory Medicine, European Institute of Oncology, Milan University School of Medicine, Milan, Italy.
    Tumour growth fraction and apoptosis in salivary gland acinic cell carcinomas: Prognostic implications of Ki-67 and bcl-2 expression and of in situ end labelling (TUNEL)1997In: The Journal of Pathology, ISSN 0031-3025, Vol. 181, no 3, p. 323-329Article in journal (Refereed)
    Abstract [en]

    bcl-2 protein and Ki-67 (MIB-1) were studied in 32 acinic cell carcinomas (ACCs), all with a minimum of 5 years' clinical follow-up. Tumour apoptosis was evaluated by TdT dUTP nick end labelling (TUNEL) and by morphological criteria. Five patients died of their disease. Patients with stage I tumours had significantly better survival compared with other stages (P<0·05). Patients with MIB-1-negative tumours had significantly better survival than patients with MIB-1-positive tumours (P=0·05). This study confirms a previous report that MIB-1 is an independent prognostic factor for survival in patients with ACC. Stage I tumours had high expression of bcl-2 protein, but there was no difference when compared with other stages. TUNEL positivity was most prevalent in stage I tumours, compared with stages II, III, and IV (P<0·05), probably indicating more apoptosis. This could imply a capacity of stage I tumours ('early tumours') for early selection of tumour cells for elimination by apoptosis. There was no significant difference between expression of bcl-2 and TUNEL, between these parameters and clinical outcome, or between any parameter and morphological subclassification. We conclude that MIB-1 has prognostic value in ACC. Clinical staging, bcl-2, and TUNEL are also potentially useful as prognostic markers.

  • 2.
    Sundelin, Kaarina
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Head and Neck Cancer: Factors Affecting Tumour Growth2007Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Head and neck cancer is the fifth most common cancer worldwide with an estimated annual global incidence of over 500 000 cases. These malignant tumours develop in the mucosal linings of the upper respiratory tract or in the salivary glands. The most common sites are in the oral cavity and larynx. Treatment modalities comprising surgery and chemoradiotherapy have improved significantly during the last 20 years, but not the long-term survival of patients. The aim of this thesis was to study the different factors affecting tumour growth in head and neck cancer that may have clinical implications in the future. Factors involving apoptosis, cell cycle activity, inflammation, and enzyme activity were of special interest.

    The results of the thesis indicate that patients with malignant salivary gland tumours having the lowest level of actively replicating cells have the best prognosis. The largest amount of replicating cells in tongue cancer specimens was found in the peripheral areas of tumour nests. Metallothionein, a protein that can hinder apoptosis, was found in excess in the same areas, whereas apoptosis activity was considerably lower. Taken together, these results indicate that the most aggressive cancer cells are found in the peripheral areas of tumours where apoptosis may be hindered.

    The expression of the death receptor Fas was higher in tongue cancer specimens than in normal mucosa. The expression of this receptor was studied further in two cell lines established from oral cancers. When a low dose of cisplatin was added to cell cultures, the Fas expression was enhanced in both cell lines and, furthermore, the Fas-induced apoptosis was increased in one of the cell lines. The results show that a common chemotherapeutic drug given in a low, less toxic dose may enhance receptor-mediated apoptosis of cancer cells.

    Malignant solid tumours are often distinguished by an increased proteolytic activity resulting in invasive growth, neo-angiogenesis, and metastases. This activity is conducted by enzymes that are secreted from tumour cells, or from normal cells in the tumour microenvironment. The regulation of enzyme secretion may be mediated by cytokines, small signalling molecules also present in cancer tissue. The results of this thesis show that two cytokines can synergistically induce enzyme secretion (matrix metalloproteinase-1 and -9) from oral cancer cells. Cytokine tumour necrosis factor-alpha and hepatocyte growth factor added alone to cell cultures strongly stimulated secretion of these enzymes. Thus, the tested cytokines, which are commonly secreted by fibroblasts and immune cells, may promote tumour growth.

    This thesis has contributed to an increased understanding of factors affecting tumour growth in head and neck cancer. The upcoming cancer therapies will be based on the increasing knowledge of these and other aberrant cellular mechanisms that may vary between different cancer forms.

    List of papers
    1. Tumour growth fraction and apoptosis in salivary gland acinic cell carcinomas: Prognostic implications of Ki-67 and bcl-2 expression and of in situ end labelling (TUNEL)
    Open this publication in new window or tab >>Tumour growth fraction and apoptosis in salivary gland acinic cell carcinomas: Prognostic implications of Ki-67 and bcl-2 expression and of in situ end labelling (TUNEL)
    Show others...
    1997 (English)In: The Journal of Pathology, ISSN 0031-3025, Vol. 181, no 3, p. 323-329Article in journal (Refereed) Published
    Abstract [en]

    bcl-2 protein and Ki-67 (MIB-1) were studied in 32 acinic cell carcinomas (ACCs), all with a minimum of 5 years' clinical follow-up. Tumour apoptosis was evaluated by TdT dUTP nick end labelling (TUNEL) and by morphological criteria. Five patients died of their disease. Patients with stage I tumours had significantly better survival compared with other stages (P<0·05). Patients with MIB-1-negative tumours had significantly better survival than patients with MIB-1-positive tumours (P=0·05). This study confirms a previous report that MIB-1 is an independent prognostic factor for survival in patients with ACC. Stage I tumours had high expression of bcl-2 protein, but there was no difference when compared with other stages. TUNEL positivity was most prevalent in stage I tumours, compared with stages II, III, and IV (P<0·05), probably indicating more apoptosis. This could imply a capacity of stage I tumours ('early tumours') for early selection of tumour cells for elimination by apoptosis. There was no significant difference between expression of bcl-2 and TUNEL, between these parameters and clinical outcome, or between any parameter and morphological subclassification. We conclude that MIB-1 has prognostic value in ACC. Clinical staging, bcl-2, and TUNEL are also potentially useful as prognostic markers.

    Keywords
    apoptosis, bcl-2, TUNEL, end labelling, salivary neoplasm, Ki-67
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-12804 (URN)10.1002/(SICI)1096-9896(199703)181:3<323::AID-PATH780>3.0.CO;2-K (DOI)
    Available from: 2007-11-28 Created: 2007-11-28 Last updated: 2009-08-18
    2. Metallothionein and Fas (CD95) are expressed in squamous cell carcinoma of the tongue
    Open this publication in new window or tab >>Metallothionein and Fas (CD95) are expressed in squamous cell carcinoma of the tongue
    Show others...
    1997 (English)In: European Journal of Cancer, ISSN 0959-8049, Vol. 33, no 11, p. 1860-1864Article in journal (Refereed) Published
    Abstract [en]

    Metallothionein (MT) is a chelator present in myoepithelial cells, whilst the Fas-receptor (APO-1, CD95) has been described primarily in human T Jurkat cells. 20 cases of carcinoma of the tongue were investigated immunocytochemically with regard to MT, Fas and Bcl-2. In normal oral squamous epithelium, MT is located in the basal/parabasal dividing cells only. In well-differentiated nests of carcinomas, MT is observed almost entirely in peripherally located cells. In situ end-labelling indicates apoptosis in the centre of these nests, but not in the peripheral areas. Less-differentiated areas show more general MT-positivity, but little apoptosis. All 24 tumours are Fas-positive, but normal epithelia are mainly negative (P< 0.0001). Bcl-2 protein was sparse in the tumours compared with MT and Fas (P< 0.0001). We thus suggest that MT, possibly due to its chelating properties, may contribute to delaying cells entering apoptosis, both in normal epithelium near the base and in less-differentiated regions of carcinoma. Moreover, Fas may be present in cells of human malignancies, as well as those of established malignant cell lines.

    Keywords
    apoptosis, Bcl-2, carcinoma squamous cell, chelator, Fas, metallothionein, tongue
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-12805 (URN)10.1016/S0959-8049(97)00216-5 (DOI)
    Available from: 2007-11-28 Created: 2007-11-28 Last updated: 2009-08-21
    3. Effects of cisplatin, interferon-alpha and 13-cis retinoic acid on the expression of Fas (CD95), intercellular adhesion molecule-1 (ICAM-1) and epidermal growth factor receptor (EGFR) in oral cancer cell lines
    Open this publication in new window or tab >>Effects of cisplatin, interferon-alpha and 13-cis retinoic acid on the expression of Fas (CD95), intercellular adhesion molecule-1 (ICAM-1) and epidermal growth factor receptor (EGFR) in oral cancer cell lines
    2007 (English)In: Journal of Oral Pathology & Medicine, ISSN 0904-2512, E-ISSN 1600-0714, Vol. 36, no 3, p. 177-183Article in journal (Refereed) Published
    Abstract [en]

    Background: Previous studies showed that many chemotherapeutic agents can induce immuno-suppression at therapeutic drug concentrations whereas low drug doses induce immuno-augmentation.

    Methods: The effect of low-dose cisplatin, interferon-alpha, and 13-cis retinoic acid on receptors involved in immune-mediated apoptosis (Fas/CD95), cell growth (epidermal growth factor receptor) and lymphocyte adhesion (intercellular adhesion molecule-1) was investigated in two oral cancer cell lines (UT-SCC-20A and UT-SCC-24A). Different methods for cell preparation were studied: mechanical and enzymatic detachment, and culture on chamber slides. Receptor expression was investigated using immunohistochemical staining. The amount of soluble and cell-bound Fas was determined with the ELISA technique, and the functional relevance of Fas expression, apoptosis induction, was analyzed.

    Results: Cisplatin enhanced cytoplasm and membrane staining for Fas in both cell lines. After cisplatin treatment, the amount of soluble Fas was increased in UT-SCC-20A cultures, but no effect was observed in the UT-SCC-24A cell line. Apoptosis, measured as enhanced caspase-3 activity, was induced by an agonistic Fas antibody (CH11) after cisplatin treatment in UT-SCC-24A cells.

    Conclusions: Low-dose cisplatin treatment enhanced Fas expression in both cell lines and increased susceptibility to apoptosis in one of them.

    Place, publisher, year, edition, pages
    Blackwell Publishing, 2007
    Keywords
    apoptosis, 13-cis retinoic acid, cisplatin, EGFR, Fas (CD95), ICAM-1, α-interferon, oral cancer
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-12806 (URN)10.1111/j.1600-0714.2006.00503.x (DOI)000244244500009 ()17305640 (PubMedID)
    Available from: 2007-11-28 Created: 2007-11-28 Last updated: 2017-12-14Bibliographically approved
    4. Effects of cytokines on matrix metalloproteinase expression in squamous cell carcinoma in vitro
    Open this publication in new window or tab >>Effects of cytokines on matrix metalloproteinase expression in squamous cell carcinoma in vitro
    2007 (English)In: Acta Oto-Laryngologica, ISSN 0001-6489, E-ISSN 1651-2251, Vol. 125, no 7, p. 765-773Article in journal (Refereed) Published
    Abstract [en]

    Objective: MMPs play an important role in enhanced intra-tumoral proteolytic activity, promoting angiogenesis and invasion by acting on extracellular matrix substances. Cytokines secreted by tumour-infiltrating immune cells, fibroblasts and tumour cells can modulate MMP expression and secretion by cancer cells. The objective of this study was to investigate the effects of IL-6, soluble IL-6 receptor (sIL-6R), HGF, TNF- and IL-8 on MMP-1, -2 and -9 expression by two oral squamous cell carcinoma cell lines (UT-SCC-20A and -24A).

    Material and methods: ELISA was used to analyse secretion of total MMP protein and gelatin zymography was used for activity analysis.

    Results: IL-6 had a moderate stimulatory effect on MMP-1 secretion in both cell lines, whereas sIL-6R had no effect. When these cytokines were added together, a dose-dependent, synergistic stimulatory effect was observed. HGF also upregulated MMP-1 secretion, especially in one cell line (UT-SCC-24A), and a synergistic effect was observed when HGF was added to IL-6 in both cell lines. MMP-9 secretion by UT-SCC-24A was increased when stimulated with HGF and IL-6 combined with sIL-6R, whereas no effect was found in the other cell line. TNF- stimulated MMP-9 secretion in both cell lines, but only stimulated MMP-1 secretion in one (UT-SCC-24A). The zymographic results were consistent with the ELISA results, indicating an upregulation of active enzyme when a stimulatory effect on protein expression was detected.

    Conclusions: The intra-tumoral cytokines IL-6, hepatocyte growth factor (HGF) and tumour necrosis factor- (TNF-) stimulate oral cancer cells to enhanced secretion of matrix metalloproteinase (MMP)-1 and -9. These results contribute to an understanding of the extracellular events necessary for tumour progression.

    Keywords
    Cytokine, matrix metalloproteinase, oral cancer, squamous cell carcinoma
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:liu:diva-12807 (URN)10.1080/00016480510027484 (DOI)
    Available from: 2007-11-28 Created: 2007-11-28 Last updated: 2017-12-14
  • 3.
    Sundelin, Kaarina
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Jadner, M.
    Norberg-Spaak, Lena
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Davidsson, A.
    Department of Otorhinolaryngology, Örebro Medical Center Hospital, Örebro, Sweden.
    Hellquist, H.B.
    Eur, J.
    Metallothionein and Fas (CD95) are expressed in squamous cell carcinoma of the tongue1997In: European Journal of Cancer, ISSN 0959-8049, Vol. 33, no 11, p. 1860-1864Article in journal (Refereed)
    Abstract [en]

    Metallothionein (MT) is a chelator present in myoepithelial cells, whilst the Fas-receptor (APO-1, CD95) has been described primarily in human T Jurkat cells. 20 cases of carcinoma of the tongue were investigated immunocytochemically with regard to MT, Fas and Bcl-2. In normal oral squamous epithelium, MT is located in the basal/parabasal dividing cells only. In well-differentiated nests of carcinomas, MT is observed almost entirely in peripherally located cells. In situ end-labelling indicates apoptosis in the centre of these nests, but not in the peripheral areas. Less-differentiated areas show more general MT-positivity, but little apoptosis. All 24 tumours are Fas-positive, but normal epithelia are mainly negative (P< 0.0001). Bcl-2 protein was sparse in the tumours compared with MT and Fas (P< 0.0001). We thus suggest that MT, possibly due to its chelating properties, may contribute to delaying cells entering apoptosis, both in normal epithelium near the base and in less-differentiated regions of carcinoma. Moreover, Fas may be present in cells of human malignancies, as well as those of established malignant cell lines.

  • 4.
    Sundelin, Kaarina
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Roberg, Karin
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Grénman, Reidar
    Department of Otorhinolaryngology – Head and Neck Surgery and Department of Medical Biochemistry, University Hospital and University of Turku, Turku, Finland.
    Håkansson, Leif
    Östergötlands Läns Landsting, Centre of Surgery and Oncology, Department of Oncology UHL.
    Effects of cisplatin, interferon-alpha and 13-cis retinoic acid on the expression of Fas (CD95), intercellular adhesion molecule-1 (ICAM-1) and epidermal growth factor receptor (EGFR) in oral cancer cell lines2007In: Journal of Oral Pathology & Medicine, ISSN 0904-2512, E-ISSN 1600-0714, Vol. 36, no 3, p. 177-183Article in journal (Refereed)
    Abstract [en]

    Background: Previous studies showed that many chemotherapeutic agents can induce immuno-suppression at therapeutic drug concentrations whereas low drug doses induce immuno-augmentation.

    Methods: The effect of low-dose cisplatin, interferon-alpha, and 13-cis retinoic acid on receptors involved in immune-mediated apoptosis (Fas/CD95), cell growth (epidermal growth factor receptor) and lymphocyte adhesion (intercellular adhesion molecule-1) was investigated in two oral cancer cell lines (UT-SCC-20A and UT-SCC-24A). Different methods for cell preparation were studied: mechanical and enzymatic detachment, and culture on chamber slides. Receptor expression was investigated using immunohistochemical staining. The amount of soluble and cell-bound Fas was determined with the ELISA technique, and the functional relevance of Fas expression, apoptosis induction, was analyzed.

    Results: Cisplatin enhanced cytoplasm and membrane staining for Fas in both cell lines. After cisplatin treatment, the amount of soluble Fas was increased in UT-SCC-20A cultures, but no effect was observed in the UT-SCC-24A cell line. Apoptosis, measured as enhanced caspase-3 activity, was induced by an agonistic Fas antibody (CH11) after cisplatin treatment in UT-SCC-24A cells.

    Conclusions: Low-dose cisplatin treatment enhanced Fas expression in both cell lines and increased susceptibility to apoptosis in one of them.

  • 5.
    Sundelin, Kaarina
    et al.
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Roberg, Karin
    Linköping University, Department of Clinical and Experimental Medicine, Oto-Rhiono-Laryngology and Head & Neck Surgery . Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of ENT - Head and Neck Surgery UHL.
    Grénman, Reidar
    Departments of Otorhinolaryngology—Head and Neck Surgery and Medical Biochemistry, University Hospital and University of Turku, Turku, Finland.
    Håkansson, Leif
    Department of Medical Oncology, University Hospital, Linköping, Sweden.
    Effects of cytokines on matrix metalloproteinase expression in squamous cell carcinoma in vitro2007In: Acta Oto-Laryngologica, ISSN 0001-6489, E-ISSN 1651-2251, Vol. 125, no 7, p. 765-773Article in journal (Refereed)
    Abstract [en]

    Objective: MMPs play an important role in enhanced intra-tumoral proteolytic activity, promoting angiogenesis and invasion by acting on extracellular matrix substances. Cytokines secreted by tumour-infiltrating immune cells, fibroblasts and tumour cells can modulate MMP expression and secretion by cancer cells. The objective of this study was to investigate the effects of IL-6, soluble IL-6 receptor (sIL-6R), HGF, TNF- and IL-8 on MMP-1, -2 and -9 expression by two oral squamous cell carcinoma cell lines (UT-SCC-20A and -24A).

    Material and methods: ELISA was used to analyse secretion of total MMP protein and gelatin zymography was used for activity analysis.

    Results: IL-6 had a moderate stimulatory effect on MMP-1 secretion in both cell lines, whereas sIL-6R had no effect. When these cytokines were added together, a dose-dependent, synergistic stimulatory effect was observed. HGF also upregulated MMP-1 secretion, especially in one cell line (UT-SCC-24A), and a synergistic effect was observed when HGF was added to IL-6 in both cell lines. MMP-9 secretion by UT-SCC-24A was increased when stimulated with HGF and IL-6 combined with sIL-6R, whereas no effect was found in the other cell line. TNF- stimulated MMP-9 secretion in both cell lines, but only stimulated MMP-1 secretion in one (UT-SCC-24A). The zymographic results were consistent with the ELISA results, indicating an upregulation of active enzyme when a stimulatory effect on protein expression was detected.

    Conclusions: The intra-tumoral cytokines IL-6, hepatocyte growth factor (HGF) and tumour necrosis factor- (TNF-) stimulate oral cancer cells to enhanced secretion of matrix metalloproteinase (MMP)-1 and -9. These results contribute to an understanding of the extracellular events necessary for tumour progression.

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