Le traitement des tumeurs cérébrales représente un défi d'envergure, puisqu'elles sont isolées du reste du corps par la barrière hémato-encéphalique. Il est donc impératif d'accroître nos connaissances sur les cellules composant ces tumeurs. L'une des hypothèses émergentes concerne la présence d'une sous-population spécifique de cellules au sein de la tumeur, les cellules souches cancéreuses (CSC), impliquées dans l'initiation et la récurrence des cancers. Les marqueurs de surfaces des CSC varient largement selon le tissu d'intérêt, mais il semble que CD133, une glycoprotéine membranaire, soit une signature commune à plusieurs CSC. S'il est courant de traiter le cancer en ciblant les cellules cancéreuses elles-mêmes, cibler le microenvironnement dans lequel évolue la tumeur est une avenue thérapeutique prometteuse. En effet, les cellules endothéliales (CE) au sein d'une tumeur sont connues pour avoir des propriétés différentes des CE normales. Ces cellules endothéliales tumorales (CET) peuvent donc constituer une cible supplémentaire pour inhiber sélectivement la croissance tumorale. À l'aide d'une lignée de médulloblastome (DAOY), nous avons identifié et évalué de nouvelles caractéristiques cellulaires et moléculaires impliquées dans la régulation du phénotype invasif associé à la formation de structures comparables aux neurosphères formées par les CSC CD133(+). De plus, nous avons apporté de nouvelles informations concernant les propriétés chimiopréventives du sulforaphane (SFN), un composé naturel retrouvé principalement dans le brocoli, en ciblant spécifiquement les CET cérébrales possiblement impliquées dans la cooption vasculaire. En utilisant la technologie de l'ARN interférant, nous avons montré que MMP-9 et MT1-MMP, deux métalloprotéinases matricielles importantes pour l'invasion cellulaire, les métastases et la résistance à la radiation, avaient un rôle crucial dans la formation par les DAOY de structures apparentées aux neurosphères CD133(+). De plus, nous avons mis en évidence une inhibition presque complète (> 90 %) de la migration des HBMEC (human brain microvascular endothelial cell) activées par un carcinogène, le phorbol 12-myristate 13-acétate (PMA). Ainsi, nos résultats suggèrent que les molécules présentes dans notre diète pourraient cibler les CET cérébrales composant la masse tumorale. En somme, nous proposons que MMP-9 constitue une cible de choix pour le traitement du cancer, que ce soit au niveau du compartiment tumoral ou vasculaire. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Cellule souche cancéreuse (CSC), Cellule endothéliale tumorale (CET), Sulforaphane (SFN), Métalloprotéinase matricielle-9 (MMP-9), Tumeur cérébrale.

Untersuchung der Reaktion kultivierter humaner Keratinozyten auf das Binden von Autoantikörpern beim bullösen Pemphigoid. Festgestellt wurde eine erhöhte Freisetzung von t-Pa, sowie eine Umverteilung und Reduktion des Zielantigens unter Calciumeinfluß. / Experiments to the reactions of cultivated keratinocytes after binding of autoantibodies against BP180. We recognizzed elevated levels of t-Pa and an alterated distribution of the target antigen under calcium influence.

Introdução: a incidência dos melanomas permanece em ascensão em diversos países. Os nevos melanocíticos podem ser seus precursores ou marcadores de risco. A radiação ultravioleta é o principal fator de risco ambiental para o seu desenvolvimento. Estudos com nevos irradiados mostram que a radiação ultravioleta B (UVB) pode causar alterações morfológicas e bioquímicas semelhantes às de um melanoma in situ. As metaloproteinases da matriz (MMP) são enzimas proteolíticas e, particularmente, as MMP-2 e –9 (gelatinases A e B) parecem estar associadas à invasão tumoral, à formação de metástases e de neoangiogênese em melanomas. O objetivo do presente estudo é avaliar os efeitos da UVB nas expressões imunoistoquímicas de MMP-2 e –9 nas diferentes linhagens celulares de nevos melanocíticos. Métodos: quarenta e dois nevos melanocíticos tiveram suas metades irradiadas com dose de 2 DEM (dose eritematosa mínima) de UVB e foram excisados uma semana após. As expressões imunoistoquímicas das MMP-2 e -9 foram comparadas, quanto à sua intensidade, por três avaliadores diferentes entre os lados irradiados e não irradiados em queratinócitos, melanócitos de epiderme e derme superior, células endoteliais e fibroblastos. Os dados foram analisados pelo teste t pareado para as diferenças de expressão e pelo ICC para avaliação da homogeneidade entre as respostas dos observadores. Resultados: com relação à expressão imunoistoquímica de MMP-2, todas as linhagens celulares mostraram aumento no lado irradiado, especialmente os melanócitos epidérmicos. Quanto à MMP-9, somente nos queratinócitos, não se observou aumento de expressão do lado irradiado, ficando essa evidente nas demais linhagens celulares avaliadas. Conclusões: A UVB na dose de 2 DEM aumenta a expressão imunoistoquímica das MMP-2 e –9 em quase todas as linhagens celulares dos nevos melanocíticos avaliados até uma semana após a irradiação, com exceção feita queratinócitos, com a MMP-9. / Background: the incidence of melanoma continues to increase in several countries. Melanocytic nevi may represent precursors or risk indicators of cutaneous melanoma. Ultraviolet radiation is the main environmental risk factor in their development. Studies with irradiated nevi show that ultraviolet B (UVB) radiation can cause morphological and biological alterations similar to those of a melanoma in situ. Matrix metalloproteinases (MMP) are proteolytic enzymes, and MMP-2 and -9 (gelatinase A and B) in particular, appear to be involved with tumour invasion, the formation of metastases and neoangiogenesis in melanomas. This study aims to evaluate the effects of UVB radiation on the immunohistochemical expression of MMP–2 and –9 in different cell lines from melanocytic nevi. Methods: one half of each of the forty-two melanocytic nevi used in the study was irradiated with 2 MEDs (Minimal Erythema Dosis) of UVB radiation and excised one week later. Three different observers were given the task of comparing the intensity of the immunohistochemical expression of the MMP –2 and –9 on the irradiated and nonirradiated sides of keratinocytes, melanocytes from the epidermis and upper dermis, endothelial cells and fibroblasts. The collected data were analysed using the paired t test for differences in expression and ICC in order to assess the homogeneity of the evaluations made by the observers. Results: in relation to the expression of MMP–2, all the cell lines showed an increase on the irradiated sides, especially the epidermal melanocytes. Regarding MMP-9, while no significant increase in its expression in keratinocytes was noted on the irradiated side, significant increases were observed in the remaining lines. Conclusions: UVB radiation at 2 MEDs increases the immunohistochemical expression of MMP –2 and –9 in almost all evaluated cell lines up to one week after irradiation, with the exception of MMP-9 in keratinocytes.

Introdução: a incidência dos melanomas permanece em ascensão em diversos países. Os nevos melanocíticos podem ser seus precursores ou marcadores de risco. A radiação ultravioleta é o principal fator de risco ambiental para o seu desenvolvimento. Estudos com nevos irradiados mostram que a radiação ultravioleta B (UVB) pode causar alterações morfológicas e bioquímicas semelhantes às de um melanoma in situ. As metaloproteinases da matriz (MMP) são enzimas proteolíticas e, particularmente, as MMP-2 e –9 (gelatinases A e B) parecem estar associadas à invasão tumoral, à formação de metástases e de neoangiogênese em melanomas. O objetivo do presente estudo é avaliar os efeitos da UVB nas expressões imunoistoquímicas de MMP-2 e –9 nas diferentes linhagens celulares de nevos melanocíticos. Métodos: quarenta e dois nevos melanocíticos tiveram suas metades irradiadas com dose de 2 DEM (dose eritematosa mínima) de UVB e foram excisados uma semana após. As expressões imunoistoquímicas das MMP-2 e -9 foram comparadas, quanto à sua intensidade, por três avaliadores diferentes entre os lados irradiados e não irradiados em queratinócitos, melanócitos de epiderme e derme superior, células endoteliais e fibroblastos. Os dados foram analisados pelo teste t pareado para as diferenças de expressão e pelo ICC para avaliação da homogeneidade entre as respostas dos observadores. Resultados: com relação à expressão imunoistoquímica de MMP-2, todas as linhagens celulares mostraram aumento no lado irradiado, especialmente os melanócitos epidérmicos. Quanto à MMP-9, somente nos queratinócitos, não se observou aumento de expressão do lado irradiado, ficando essa evidente nas demais linhagens celulares avaliadas. Conclusões: A UVB na dose de 2 DEM aumenta a expressão imunoistoquímica das MMP-2 e –9 em quase todas as linhagens celulares dos nevos melanocíticos avaliados até uma semana após a irradiação, com exceção feita queratinócitos, com a MMP-9. / Background: the incidence of melanoma continues to increase in several countries. Melanocytic nevi may represent precursors or risk indicators of cutaneous melanoma. Ultraviolet radiation is the main environmental risk factor in their development. Studies with irradiated nevi show that ultraviolet B (UVB) radiation can cause morphological and biological alterations similar to those of a melanoma in situ. Matrix metalloproteinases (MMP) are proteolytic enzymes, and MMP-2 and -9 (gelatinase A and B) in particular, appear to be involved with tumour invasion, the formation of metastases and neoangiogenesis in melanomas. This study aims to evaluate the effects of UVB radiation on the immunohistochemical expression of MMP–2 and –9 in different cell lines from melanocytic nevi. Methods: one half of each of the forty-two melanocytic nevi used in the study was irradiated with 2 MEDs (Minimal Erythema Dosis) of UVB radiation and excised one week later. Three different observers were given the task of comparing the intensity of the immunohistochemical expression of the MMP –2 and –9 on the irradiated and nonirradiated sides of keratinocytes, melanocytes from the epidermis and upper dermis, endothelial cells and fibroblasts. The collected data were analysed using the paired t test for differences in expression and ICC in order to assess the homogeneity of the evaluations made by the observers. Results: in relation to the expression of MMP–2, all the cell lines showed an increase on the irradiated sides, especially the epidermal melanocytes. Regarding MMP-9, while no significant increase in its expression in keratinocytes was noted on the irradiated side, significant increases were observed in the remaining lines. Conclusions: UVB radiation at 2 MEDs increases the immunohistochemical expression of MMP –2 and –9 in almost all evaluated cell lines up to one week after irradiation, with the exception of MMP-9 in keratinocytes.

Introdução: a incidência dos melanomas permanece em ascensão em diversos países. Os nevos melanocíticos podem ser seus precursores ou marcadores de risco. A radiação ultravioleta é o principal fator de risco ambiental para o seu desenvolvimento. Estudos com nevos irradiados mostram que a radiação ultravioleta B (UVB) pode causar alterações morfológicas e bioquímicas semelhantes às de um melanoma in situ. As metaloproteinases da matriz (MMP) são enzimas proteolíticas e, particularmente, as MMP-2 e –9 (gelatinases A e B) parecem estar associadas à invasão tumoral, à formação de metástases e de neoangiogênese em melanomas. O objetivo do presente estudo é avaliar os efeitos da UVB nas expressões imunoistoquímicas de MMP-2 e –9 nas diferentes linhagens celulares de nevos melanocíticos. Métodos: quarenta e dois nevos melanocíticos tiveram suas metades irradiadas com dose de 2 DEM (dose eritematosa mínima) de UVB e foram excisados uma semana após. As expressões imunoistoquímicas das MMP-2 e -9 foram comparadas, quanto à sua intensidade, por três avaliadores diferentes entre os lados irradiados e não irradiados em queratinócitos, melanócitos de epiderme e derme superior, células endoteliais e fibroblastos. Os dados foram analisados pelo teste t pareado para as diferenças de expressão e pelo ICC para avaliação da homogeneidade entre as respostas dos observadores. Resultados: com relação à expressão imunoistoquímica de MMP-2, todas as linhagens celulares mostraram aumento no lado irradiado, especialmente os melanócitos epidérmicos. Quanto à MMP-9, somente nos queratinócitos, não se observou aumento de expressão do lado irradiado, ficando essa evidente nas demais linhagens celulares avaliadas. Conclusões: A UVB na dose de 2 DEM aumenta a expressão imunoistoquímica das MMP-2 e –9 em quase todas as linhagens celulares dos nevos melanocíticos avaliados até uma semana após a irradiação, com exceção feita queratinócitos, com a MMP-9. / Background: the incidence of melanoma continues to increase in several countries. Melanocytic nevi may represent precursors or risk indicators of cutaneous melanoma. Ultraviolet radiation is the main environmental risk factor in their development. Studies with irradiated nevi show that ultraviolet B (UVB) radiation can cause morphological and biological alterations similar to those of a melanoma in situ. Matrix metalloproteinases (MMP) are proteolytic enzymes, and MMP-2 and -9 (gelatinase A and B) in particular, appear to be involved with tumour invasion, the formation of metastases and neoangiogenesis in melanomas. This study aims to evaluate the effects of UVB radiation on the immunohistochemical expression of MMP–2 and –9 in different cell lines from melanocytic nevi. Methods: one half of each of the forty-two melanocytic nevi used in the study was irradiated with 2 MEDs (Minimal Erythema Dosis) of UVB radiation and excised one week later. Three different observers were given the task of comparing the intensity of the immunohistochemical expression of the MMP –2 and –9 on the irradiated and nonirradiated sides of keratinocytes, melanocytes from the epidermis and upper dermis, endothelial cells and fibroblasts. The collected data were analysed using the paired t test for differences in expression and ICC in order to assess the homogeneity of the evaluations made by the observers. Results: in relation to the expression of MMP–2, all the cell lines showed an increase on the irradiated sides, especially the epidermal melanocytes. Regarding MMP-9, while no significant increase in its expression in keratinocytes was noted on the irradiated side, significant increases were observed in the remaining lines. Conclusions: UVB radiation at 2 MEDs increases the immunohistochemical expression of MMP –2 and –9 in almost all evaluated cell lines up to one week after irradiation, with the exception of MMP-9 in keratinocytes.

Orientador: José Mauro Granjeiro / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-09T02:30:54Z (GMT). No. of bitstreams: 1 Oliveira_RodrigoCardosode_D.pdf: 3052048 bytes, checksum: f37f738c374af69b889b7672904eddfc (MD5) Previous issue date: 2005 / Resumo: O objetivo do trabalho foi avaliar comparativamente a neoformação óssea e o perfil de gelatinases 2 e 9 durante o reparo de defeito crítico em crânio de ratos tratados com osso autógeno ou xenoenxerto desmineralizado. Um defeito ósseo de tamanho crítico (8 mm) foi confeccionado no crânio de 90 ratos Wistar (90 dias de vida), e preenchido com osso autógeno (grupo controle) obtido durante a confecção do defeito ou com matriz óssea bovina desmineralizada (grupo teste). Após os períodos de 7, 14, 21, 30 e 90 dias, os animais foram eutanasiados e as peças coletadas para análises histomorfométrica (em hematoxilina e eosina) e zimográfica. A análise paramétrica foi realizada utilizando análise de variância (teste de Tukey se p<0,05). O completo fechamento do defeito no grupo controle foi observado aos 90 dias com a neoformação óssea ocorrendo das bordas do defeito para o centro e da dura-máter para epiderme. No grupo teste houve atraso no processo de reparo, ossificação incompleta e substituição das partículas do biomaterial por tecido conjuntivo fibroso, após 21 dias. Nos primeiros 14 dias após a cirurgia o infiltrado inflamatório predominante era composto de células mononucleares e poucas células gigantes multinucleadas. A análise zimográfica demonstrou que a atividade MMP-2 e -9 foram significativamente maiores no grupo teste que no grupo controle (p<0,05), sendo que a atividade MMP-2 manteve-se elevada até o período de 14 dias no grupo teste. A despeito da biocompatibilidade do xenoenxerto, o biomaterial não foi capaz de promover a neoformação óssea no defeito, possivelmente devido ao intenso estímulo da atividade gelatinolítica, em particular da MMP-2, que pode ter mediado a reabsorção prematura da matriz óssea bovina desmineralizada / Abstract: The purpose of this study was to evaluate comparatively new bone formation and the profile of the gelatinases 2 and 9 during the repair of critical size defects treated with autogenous bone or demineralized xenograft. A critical defect (8mm) was created in the skull of 90 Wistar rats (90-day-old) and treated with autogenous bone (control group) obtained during the confection of the defect or demineralized bovine bone (experimental group). After at 7, 14, 21, 30 and 90 days, the animals were killed and the calvaria removed for morphometric (stained by hematoxylin-eosin) and zymografic analysis. Parametric analysis was realized with analysis of variance (Tuckey¿s test if p<0.05). The control group showed complete closure of the defects at 90 days with new bone formation occurring from the sides towards the center of the defect and from the dura-mater outwards to the epidermis. In the experimental group, there was a delay in the process of repair, incomplete ossification and nearly complete substitution of material particles by fibrotic connective tissue after 21 days. At 14 days post-operatively, the inflammatory infiltrate consisted predominantly of mononuclear cells and few multinuclear giants cells. Zymografic analysis demonstrated that the activities of MMP-2 and -9 were significantly higher in the experimental group than in the control group (p<0.05), in addition the activity of MMP-2 was increased up to 14 days in control group . Despite the biocompatibity of the xenograft, the biomaterial was not capable to promote new bone formation in the defect. This might be possibly related to the intense stimulation of the gelanolitic activity, in particular of MMP-2, which in turn may have mediated the resorption of the demineralized bovine bone / Doutorado / Bioquimica / Doutor em Biologia Funcional e Molecular

Abstract The invasion of a tumour through tissue limiting basement membranes is the critical step in malignant growth. Gelatinases (MMP-2 and MMP-9) are endopeptidases capable of degrading extracellular and pericellular matrix proteins such as collagen IV, the major component of basement membranes. An over-expression of these gelatinases is generally found in malignant tumours and is linked to impaired prognosis in many cancer types. Tissue inhibitors of metalloproteinases (TIMPs), endogenous regulators of the MMP activity, have recently been introduced as multifunctional proteins, which have paradoxical roles in tumour growth. Little data exists on the clinical significance of the gelatinases and TIMPs in gynaecological cancers. In this study the clinical significance of the gelatinases was studied in endometrial and uterine cervical cancers by using immunohistochemical staining with specific antibodies. In epithelial ovarian cancer (EOC) these enzymes and their TIMPs were studied in the preoperative serum samples using ELISA assay. Additionally, sequential serum measurements were performed during chemotherapy to evaluate them as treatment response indicators. In endometrial cancer, MMP-9 positivity correlated to a poor histological differentiation and an advanced clinical stage. High MMP-2 expression correlated to a poor differentiation, and unfavourable survival in stage I cancers, with mortality rates of 5% and 19% in patients with MMP-2 negative versus intensively MMP-2 positive tumours, respectively. In cervical cancers high MMP-2 expression correlated to an increased mortality risk. High MMP-9 expression was connected to a good differentiation of a tumour. In EOC, a high circulating TIMP-1 value correlated to all the examined aggressive features of EOC, including poor survival. The serum measurements of TIMP-1 were uninformative about response evaluation during chemotherapy but paradoxically, an increase in gelatinases and TIMP-2 seemed to reflect a good response to treatment. In conclusion, the data from this study show that high MMP-2 expression in tumour tissue could be prognostic in endometrial and cervical cancer, and preoperative circulating TIMP-1 could serve as an additional prognostic marker in EOC. Studies with larger patient cohorts would be necessary to further explore the value of these enzymes in clinical practice in gynaecological cancers.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

Εισαγωγή: Ο καρκίνος του λάρυγγα, ιδιαιτέρως σε προχωρημένα στάδια, είναι μία καταστροφική νόσος η οποία χαρακτηρίζεται από αυξημένη διηθητικότητα και μεταστατικότητα. Η ανεύρεση ενός δείκτη πρώιμης διάγνωσης, παρακολούθησης και πρόγνωσης της νόσου θα ήταν ιδιαίτερα ευπρόσδεκτη. Συνεχώς αυξανόμενα δεδομένα στη βιβλιογραφία υποστηρίζουν την προγνωστική αξία των ζελατινασών και τον πιθανό ρόλο τους ως μοριακών δεικτών μεταξύ άλλων και στον καρκίνο του λάρυγγα. Σκοπός: Η διαπίστωση μεταβολών στα επίπεδα ορού των ζελατινασών Α και Β σε ασθενείς με καρκίνο του λάρυγγα μετά από εφαρμογή θεραπείας, καθώς και η πιθανή συσχέτιση με διάφορες κλινικοπαθολογικές παραμέτρους πριν και μετά τη θεραπευτική παρέμβαση. Υλικό και μέθοδος: Σαράντα εννέα ασθενείς και 8 υγιείς μάρτυρες συμπεριλήφθηκαν στη μελέτη. Ελήφθησαν προεγχειρητικά και μετεγχειρητικά δείγματα ορού τα οποία στη συνέχεια υποβλήθηκαν σε ζυμογραφία ζελατίνης. Η παρουσία ζελατινασών επιβεβαιώθηκε με την τεχνική western blotting. Οι ζώνες λύσης ποσοτικοποιήθηκαν με τη χρήση Scion Image PC. Η ανάλυση των αποτελεσμάτων πραγματοποιήθηκε με το πρόγραμμα SPSS 17 (SPSS Inc, Chicago, IL, USA). Αποτελέσματα: Στα ζυμογραφήματα αποτυπώθηκαν μόνο οι λανθάνουσες μορφές των ενζύμων (προένζυμα). Τα προ της θεραπείας επίπεδα και των δύο ζελατινασών στον ορό του αίματος των ασθενών με καρκίνο του λάρυγγα ήταν σημαντικά υψηλότερα σε σχέση με αυτά των υγιών μαρτύρων. Ασθενείς με υπεργλωττιδικό καρκίνωμα και ενεργοί καπνιστές είχαν σημαντικά υψηλότερα επίπεδα proMMP-2 σε σχέση με ασθενείς που έπασχαν από γλωττιδικό καρκίνωμα και με πρώην καπνιστές αντίστοιχα. Ασθενείς με πρωτοδιαγνωσμένη νόσο και ασθενείς με λεμφαδενικές μεταστάσεις είχαν σημαντικά χαμηλότερα προ της θεραπείας επίπεδα proMMP-9 σε σχέση με ασθενείς που προσήλθαν με υποτροπή και με ασθενείς στους οποίους δεν διαπιστώθηκε επιχώρια νόσος αντίστοιχα. Κατά τη διάρκεια της συστηματικής παρακολούθησης τα επίπεδα της proMMP-2 στον ορό παρουσίασαν σημαντική αύξηση τις πρώτες 10 με 15 ημέρες μετά την εφαρμογή θεραπείας, για να μειωθούν σταδιακά εντός των επόμενων μηνών. Οι ενεργοί καπνιστές παρουσίασαν σημαντική μείωση των επιπέδων της proMMP-2 κατά την περίοδο παρακολούθησης, σε αντίθεση με τους πρώην καπνιστές οι οποίοι εμφάνισαν σημαντική αύξηση κατά το ίδιο χρονικό διάστημα. Οι ασθενείς σταδίου ΙΙ είχαν σημαντικά χαμηλότερα επίπεδα proMMP-2 σε σχέση με ασθενείς προχωρημένων σταδίων πέντε με οκτώ μήνες μετά τη θεραπεία, όπως και οι ασθενείς οι οποίοι υποβλήθηκαν σε συντηρητική αντιμετώπιση σε σχέση με τους χειρουργημένους ασθενείς. Τα επίπεδα της proMMP-9 στον ορό επίσης παρουσίασαν σημαντική πτώση μετά την εφαρμογή θεραπείας. Διαφορές στο ρυθμό μείωσης των επιπέδων της proMMP-9 παρατηρήθηκαν μεταξύ των διαφόρων ομάδων ως προς το στάδιο, τη διαφοροποίηση, την εντόπιση, τον τύπο της νόσου (πρωτοδιαγνωσμένη ή υποτροπή), τις λεμφαδενικές μεταστάσεις, τον τρόπο αντιμετώπισης και την κατανάλωση αλκοόλ. Ωστόσο αυτή η διαφορά δεν διατηρήθηκε πέντε με οκτώ μήνες μετά την εφαρμογή θεραπείας, με εξαίρεση την ομάδα των χειρουργημένων ασθενών, οι οποίοι διατήρησαν σημαντικά υψηλότερα επίπεδα ενζύμου στον ορό. Αύξηση των ζελατινασών παρατηρήθηκε στον ορό ασθενών που εκδήλωσαν υποτροπή μετά από αντιμετώπιση πρωτοδιαγνωσμένης νόσου σε σχέση με αυτούς που δεν υποτροπίασαν. Ωστόσο εξαιτίας του μικρού δείγματος δεν είναι δυνατόν να εξαχθούν ασφαλή συμπεράσματα. Συμπεράσματα: Αν και δεν υφίστανται φυσιολογικές τιμές, το πρότυπο μεταβολής των επιπέδων της proMMP-9 στον ορό μετά από θεραπεία καταδεικνύει πιθανές ιδιότητες μοριακού δείκτη. Ωστόσο υπάρχουν ενδείξεις ότι και οι δύο ζελατινάσες θα μπορούσαν να χρησιμοποιηθούν για την εξατομικευμένη παρακολούθηση ασθενών με καρκίνο του λάρυγγα. Περαιτέρω έρευνα απαιτείται για την αποσαφήνιση του ζητήματος. / Introduction: Laryngeal cancer, especially in the advanced stages, is a highly devastating disease, characterized by increased invasiveness and high rates of metastasis. The identification of reliable tumour marker for prompt diagnosis, surveillance and prognosis would be highly desirable. There is a growing body of evidence with regard to the prognostic value of gelatinases and their possible role as tumour markers. Aim: To identify the pattern of alteration of serum gelatinases A and B in patients with laryngeal cancer following treatment, and a possible correlation with various clinicopathological parameters prior to and past treatment. Materials and methods: Forty nine patients and 8 healthy controls were included in the study. Pre-treatment and post-treatment serum samples were collected and processed by gelatin zymography. The presence of gelatinases was verified by western blotting. The zymograms were scanned by a digital scanner and the lysis bands were quantified by Scion Image PC. Analysis of the quantitative results was performed by using SPSS 17 (SPSS Inc, Chicago, IL, USA). Results: Only the latent forms of MMP-2 and -9 (proforms) were identified. Both gelatinases were increased in the serum of laryngeal cancer patients compared to healthy individuals. Patients with supraglottic tumours and active smokers had significantly higher pre-treatment levels of proMMP-2 than patients with glottic tumours and ex-smokers, respectively. Patients with primary disease and patients with lymph node involvement showed lower proMMP-9 pre-treatment levels than patients with recurrence and patients without neck disease, respectively. During the follow-up period the proMMP-2 serum levels increased significantly in the first ten to fifteen days after treatment, gradually decreasing over the following months. Smokers showed a very high decrease rate of proMMP-2 levels during the follow-up period, whereas in ex-smokers proMMP-2 levels significantly increased. Stage II patients showed significantly lower levels of circulating enzyme compared to patients with more advanced disease five to eight months past treatment. Similarly, conservative management was associated with lower levels of serum proMMP-2 compared to surgical management five to eight months following treatment. The proMMP-9 serum levels also showed a gradual decrease after treatment, which was statistically significant. Significant alterations in the rate of decrease developed among groups with regard to stage, grade, location, type of disease (primary or recurrence), regional disease, treatment modality and alcohol consumption. Nevertheless those differences were not maintained five to eight months past treatment, with the exception of patients who underwent surgery and who maintained higher levels of proMMP-9. An increase to the levels of both gelatinases were observed in patients with recurrent disease after having been treated for a primary compared to patients who did not develop a recurrence. However, the small sample of patients with recurrent disease during the follow-up period does not allow extrapolating sound conclusions. Conclusions: Although as yet normal values have not been established in the literature, the post-treatment alteration pattern of proMMP-9 serum levels indicates that this enzyme might play a role as a tumour marker. Nevertheless this study provides evidence that both gelatinases might be useful for surveillance on strictly individual basis in laryngeal cancer patients. Further research is necessary to clarify the contribution of both gelatinases to the disease progress and determine their role as prognostic factors and tumour markers.