Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Η έκφραση των MMP και TIMP γονιδίων και η ενεργοποίηση των MMPs έχει συσχετιστεί με την εξέλιξη του καρκίνου του μαστού και η υπερέκφραση αυτών σχετίζεται με φτωχή πρόγνωση για τον ασθενή (Bjorklund et al 2005, Wurtz et al 2005). Σε αυτή την εργασία δείχτηκε με RT-PCR ανάλυση ότι τα MCF-7 κύτταρα εκφράζουν MMP-9, MT1- και MT2-MMP, αποτελέσματα που επιβεβαιώνουν προηγούμενες εργασίες (Kousidou at al 2004, Kousidou et al 2005) και που μπορεί να σχετίζονται με τη χαμηλή δυνατότητα μετάστασης της καρκινικής αυτής σειράς. Εντούτοις MMP2-mRNA δεν ανιχνεύτηκε, παρά το γεγονός ότι η MMP2 συχνά εμφανίζεται σε κακοήθεις ιστούς στο μαστό. Η επώαση των κυττάρων με λεουπρολίδιο για 48h οδήγησε σε χαμηλότερα επίπεδα της ΜΜΡ-9 mRNA. Έχει δεiχτεί ότι, σε αντίθεση με την ΜΜΡ-2, η έκφραση της ΜΜΡ-9 εξαρτάται από την παρουσία αυξητικών παραγόντων, χημοκινών και άλλων μορίων σηματοδότησης (Bjorklund et al, 2005). Σε αυτό το πλαίσιο η υπερέκφραση της ΜΜΡ-9 δεν διατηρείται όταν τα καρκινικά κύτταρα επωάζονται σε περιβάλλον ελεύθερο τέτοιων παραγόντων, υποθέτοντας ότι η παρουσία στον ορό μερικών αυξητικών παραγόντων παρακινούν την έκφραση βασικών MMPs σε φυσιολογικά και καρκινικά κύτταρα. Από τις μεμβρανικές μεταλλοπρωτεϊνάσες οι ΜΤ1- (ΜΜΡ-14) και ΜΤ2 (ΜΜΡ-15) εμφανίζουν παρόμοια συμπεριφορά και αυξάνονται παρουσία του αγωνιστή LHRH, λεουπρολιδίου. Επιπλέον η ΜΤ1-ΜΜΡ και η ΤΙΜΡ-2 συμμετέχουν στην κύρια οδό ενεργοποίησης του ΜΜΡ-2 στην επιφάνεια των κυττάρων με τη δημιουργία ενός τριμοριακού συμπλέγματος ΜΜΡ-2, ΜΤ1-ΜΜΡ και ΤΙΜΡ-2 (Deryugina et al 2001, Lafleur et al 2003). Σε αυτήν την εργασία τα επίπεδα ΤΙΜΡ-2 mRNA επίσης αυξήθηκαν. Έτσι θα μπορούσε να ειπωθεί ότι τα αυξημένα επίπεδα ΜΤ1-ΜΜΡ και ΤΙΜΡ-2 mRNA εκφράζουν την διάθεση των κυττάρων να ενεργοποιήσουν την proMMP-2. Τα αποτελέσματα μας δείχνουν ότι τα γονίδια ΤΙΜΡ-1 και ΤΙΜΡ-3 μειώνονται παρουσία του αγωνιστή LHRH, λεουπρολιδίου. Η αύξηση των ΤΙΜΡs από τα καρκινικά κύτταρα για να εξισοροπήσουν την υψηλή προτεολυτική δράση των MMPs αποτελεί μία πιθανή θεώρηση (Brown et al, 1998). Αυτό θα μπορούσε να θεωρηθεί ως ένας αμυντικός μηχανισμός του ανθρώπινου οργανισμού για να αντιμετωπήσει την ανώμαλη αύξηση των MMPs. Έτσι τα χαμηλότερα επίπεδα του ΤΙΜΡ-1 mRNA μπορεί να σχετίζονται με τα χαμηλότερα επίπεδα ΜΜΡ-9, παρουσία του λεουπρολιδίου, αφού ο ΤΙΜΡ-1 αναστέλλει την ΜΜΡ-9 με μεγάλη εξειδίκευση. Εντούτοις ιστοί με μεγάλη περιεκτικότητα σε ΤΙΜΡ-1 mRNA και πρωτείνες σχετίζονται με κακή πρόγνωση ασθενών με καρκίνο του μαστού, παρά την ανασταλτική δράση των ΜΜΡs. Η πρόσφατη ανακάλυψη της αντι αποπτωτικής και προ αγγειογενετικής δράσης του ΤΙΜΡ-1 μπορεί να είναι μέρος αυτής της υπόθεσης (Wurtz et al, 2005). Έχοντας αυτό υπόψην τα παρατηρούμενα χαμηλά επίπεδα ΤΙΜΡ-1 mRNA μπορεί να είναι αποτέλεσμα και άλλων δράσεων με ωφέλιμο ρόλο στον καρκίνο του μαστού. Γενικά η διαφορετική έκφραση των ΤΙΜΡs στον καρκίνο του μαστού αποτελεί ένα πολύ σημαντικό θέμα, που χρήζει περαιτέρω διερεύνησης. Συμπερασματικά τα αποτελέσματά μας δεικνύουν ότι ο αγωνιστής LHRH, λεουπρολίδιο, μπορεί να τροποποιήσει σημαντικά την έκφραση των γονιδίων ΜΜΡ/ΤΙΜΡ στα κύτταρα MCF-7. Οι MMPs και ΤΙΜΡs είναι πολυδύναμα μόρια με σημαντική επίδραση στην ανάπτυξη του καρκίνου, της μετάστασης και της αγγειογένεσης. Δεδομένου ότι τα ανάλογα LHRH αποτελούν μέρος της θεραπείας ασθενών με προ ή μετά εμμηνοπαυσιακό καρκίνο του μαστού, η επίδραση των παραπάνω αναφερθέντων μεταβολών από τη χρήση του λεουρπολιδίου, χρήζει περαιτέρω διερεύνησης. / Luteinizing hormone-releasing hormone (LHRH) is not only produced by hypothalamus but also by other normal and cancer tissues. LHRH peptide agonists and antagonists inhibit the proliferation of breast cancer cells, but their effect on the expression of metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) has not been studied despite the fact that growth and invasiveness of breast cancer cells in adjacent and distant sites is associated with the expression of MMPs. In the present study, the effects of [D-Leu6,Pro9 -NHEt]LHRH, leuprolide, on gene expression of MMPs and TIMPs in the breast cancer cell line MCF-7 were examined with semi-quantitative RT-PCR. Results showed that incubation of MCF-7 with 30μM of the leuprolide for 48h in serum-containing medium resulted in a down-regulation of MMP-9 expression and increase in MT1- and MT-2 MMP mRNA levels. Furthermore, leuprolide induced a significant decrease in TIMP-1 and TIMP-3 mRNA levels and increase in TIMP-2 mRNA levels. The impact of the observed changes on the expression of MMPs and TIMPs warrants further investigation on the effects of LHRH analogues on the invasiveness and metastatic potential of breast cancer cells.

INTRODUÇÃO: Aventa-se a hipótese de quepacientes com endometriose poderiam apresentar risco elevado para doenças cardiovasculares. Existe, porém, controvérsia quanto ao perfil lipídico observado nessas pacientes. OBJETIVOS: avaliar os marcadores de risco cardiovascular associados à endometriose, comparando-se o efeito sobre eles de dois diferentes tratamentos para esta doença: Sistema Intra-uterino liberador de levonorgestrel (SIU-LNG) e o análogo do GnRH na forma de depósito (aGnRH). Marcadores inflamatórios, parâmetros clínicos e avaliação lipídica foram utilizados como marcadores de risco cardiovascular. MATERIAL E MÉTODOS: Quarenta pacientes entre 18 e 40 anos, com diagnóstico laparoscópico de endometriose, foram randomizadas para receber tratamento com SIU-LNG (n=22) e com o aGnRH (n=18), durante 6 meses. Foram avaliados o índice de massa corporal, a freqüência cardíaca, as pressões arteriais sistólica e diastólica, além dos seguintes parâmetros laboratoriais: lipidograma (colesterol total (CT), HDL-colesterol (HDL-C), LDL-colesterol (LDL-C), triglicérides (TGL)), interleucina-6 (IL-6), proteína C reativa (PCR), homocisteína, molécula de adesão decélula vascular (VCAM), fator de necrose tumoral ?(TNF-?) e contagem de leucócitos (LCT), sendo realizados no início e após seis meses de tratamento. RESULTADOS: no grupo do SIU-LNG, houve redução dos níveis do VCAM (92,8 ± 4,2ng/mL para 91,2 ± 2,7ng/mL, p=0,04), PCR (0,38 ± 0,30mg/dL para 0,28 ± 0,21mg/dL, p=0,03), CT (247,0 ± 85,0 mg/dL para 180,0 ± 31,0 mg/dL, p=0,0002), TGL (118,0 ± 76,0 mg/dL para 86,5 ± 41,5 mg/dL,p=0,003), LDL-C (160,5 + 66,0mg/dL para 114,5 + 25,5mg/dL, p=0,0005) e HDL-C (63,0 + 20,5mg/dL para 48,5 + 10,5mg/Dl, p=0,002). No grupo do aGnRH, houve aumento da homocisteína (11,5 + 2,9 µmol/L para 13,0 + 2,7µmol/L, p=0,04) e diminuição dos níveis de IL-6 (4,3 + 3,9pg/mL para 2,3 + 0,8pg/mL, p=0,005), VCAM (94,0 + 3,8ng/mL para 92,0 + 1,6ng/mL, p=0,03) e LCT (7330 + 2554 para 6350 + 1778, p=0,01). Esse estudo mostra que alguns marcadores de risco cardiovascular são influenciados por ambos aGnRH e SIU-LNG, mas esse último reduz mais os níveis lipídicos e pode ter efeitos mais favoráveis em longo prazo. / The aim of this prospective and controlled study was to evaluate the cardiovascular risk markers associated with endometriosis and the influence of the levonorgestrel intra-uterine system (LNG-IUS) compared with the GnRH analogue (GnRHa) leuprolide acetate on these risks after six monthsof treatment. Methods: This was a randomized, prospective, open clinical study, with44 patients with laparoscopically and histologically confirmed endometriosis. Patients were randomized into two groups: LNG-IUS group, 22 patients submittedto LNG-IUS insertion, and GnRHa group, 22 patients who received a monthly GnRHa injection for six months. Body mass index, systolic and diastolic arterial blood pressure, heart rate and laboratory cardiovascular risk markers such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), C-reactive protein (CRP), homocysteine (HMC), lipid profile, total leucocytes, and vascular cell adhesion molecule (VCAM) were measured before and six months after treatment. Results: in the LNG-IUS group, there was reduction of the levels of VCAM(92.8 +4.2 to 91.2+2.7 ng/mL, p = 0.04), CRP (0.38+0.30 to 0.28+0.21 mg/dL, p = 0.03), total cholesterol (247.0+85.0 to 180.0+31.0 mg/dL, p = 0.0002), triglycerides (118.0+ 76.0 to 86.5+41.5 mg/dL, p = 0.003), LDL (160.5+66.0 to 114.5+25.5 mg/dL, p = 0.0005) and HDL (63.0+20.5 to 48.5+10.5 mg/dL, p = 0.002). The GnRHa group showed an increase of HMC levels (11.5+2.9 to 13.0+2.7 µmol/L, p = 0.04) and a reduction of IL-6 levels (4.3+3.9 to 2.3+0.8 pg/mL, p = 0.005), VCAM (94.0+3.8 to 92.0+1.6 ng/mL, p = 0.03) and total leucocytes (7330+2554 to 6350+1778, p = 0.01). Conclusions: This study shows that some cardiovascular risk markers are influenced by both GnRHa and LNG-IUS,but the latter had a greater positive impact on the lipid profile, which could lead to a favorable effect during long-term treatment.

INTRODUÇÃO: Aventa-se a hipótese de quepacientes com endometriose poderiam apresentar risco elevado para doenças cardiovasculares. Existe, porém, controvérsia quanto ao perfil lipídico observado nessas pacientes. OBJETIVOS: avaliar os marcadores de risco cardiovascular associados à endometriose, comparando-se o efeito sobre eles de dois diferentes tratamentos para esta doença: Sistema Intra-uterino liberador de levonorgestrel (SIU-LNG) e o análogo do GnRH na forma de depósito (aGnRH). Marcadores inflamatórios, parâmetros clínicos e avaliação lipídica foram utilizados como marcadores de risco cardiovascular. MATERIAL E MÉTODOS: Quarenta pacientes entre 18 e 40 anos, com diagnóstico laparoscópico de endometriose, foram randomizadas para receber tratamento com SIU-LNG (n=22) e com o aGnRH (n=18), durante 6 meses. Foram avaliados o índice de massa corporal, a freqüência cardíaca, as pressões arteriais sistólica e diastólica, além dos seguintes parâmetros laboratoriais: lipidograma (colesterol total (CT), HDL-colesterol (HDL-C), LDL-colesterol (LDL-C), triglicérides (TGL)), interleucina-6 (IL-6), proteína C reativa (PCR), homocisteína, molécula de adesão decélula vascular (VCAM), fator de necrose tumoral ?(TNF-?) e contagem de leucócitos (LCT), sendo realizados no início e após seis meses de tratamento. RESULTADOS: no grupo do SIU-LNG, houve redução dos níveis do VCAM (92,8 ± 4,2ng/mL para 91,2 ± 2,7ng/mL, p=0,04), PCR (0,38 ± 0,30mg/dL para 0,28 ± 0,21mg/dL, p=0,03), CT (247,0 ± 85,0 mg/dL para 180,0 ± 31,0 mg/dL, p=0,0002), TGL (118,0 ± 76,0 mg/dL para 86,5 ± 41,5 mg/dL,p=0,003), LDL-C (160,5 + 66,0mg/dL para 114,5 + 25,5mg/dL, p=0,0005) e HDL-C (63,0 + 20,5mg/dL para 48,5 + 10,5mg/Dl, p=0,002). No grupo do aGnRH, houve aumento da homocisteína (11,5 + 2,9 µmol/L para 13,0 + 2,7µmol/L, p=0,04) e diminuição dos níveis de IL-6 (4,3 + 3,9pg/mL para 2,3 + 0,8pg/mL, p=0,005), VCAM (94,0 + 3,8ng/mL para 92,0 + 1,6ng/mL, p=0,03) e LCT (7330 + 2554 para 6350 + 1778, p=0,01). Esse estudo mostra que alguns marcadores de risco cardiovascular são influenciados por ambos aGnRH e SIU-LNG, mas esse último reduz mais os níveis lipídicos e pode ter efeitos mais favoráveis em longo prazo. / The aim of this prospective and controlled study was to evaluate the cardiovascular risk markers associated with endometriosis and the influence of the levonorgestrel intra-uterine system (LNG-IUS) compared with the GnRH analogue (GnRHa) leuprolide acetate on these risks after six monthsof treatment. Methods: This was a randomized, prospective, open clinical study, with44 patients with laparoscopically and histologically confirmed endometriosis. Patients were randomized into two groups: LNG-IUS group, 22 patients submittedto LNG-IUS insertion, and GnRHa group, 22 patients who received a monthly GnRHa injection for six months. Body mass index, systolic and diastolic arterial blood pressure, heart rate and laboratory cardiovascular risk markers such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), C-reactive protein (CRP), homocysteine (HMC), lipid profile, total leucocytes, and vascular cell adhesion molecule (VCAM) were measured before and six months after treatment. Results: in the LNG-IUS group, there was reduction of the levels of VCAM(92.8 +4.2 to 91.2+2.7 ng/mL, p = 0.04), CRP (0.38+0.30 to 0.28+0.21 mg/dL, p = 0.03), total cholesterol (247.0+85.0 to 180.0+31.0 mg/dL, p = 0.0002), triglycerides (118.0+ 76.0 to 86.5+41.5 mg/dL, p = 0.003), LDL (160.5+66.0 to 114.5+25.5 mg/dL, p = 0.0005) and HDL (63.0+20.5 to 48.5+10.5 mg/dL, p = 0.002). The GnRHa group showed an increase of HMC levels (11.5+2.9 to 13.0+2.7 µmol/L, p = 0.04) and a reduction of IL-6 levels (4.3+3.9 to 2.3+0.8 pg/mL, p = 0.005), VCAM (94.0+3.8 to 92.0+1.6 ng/mL, p = 0.03) and total leucocytes (7330+2554 to 6350+1778, p = 0.01). Conclusions: This study shows that some cardiovascular risk markers are influenced by both GnRHa and LNG-IUS,but the latter had a greater positive impact on the lipid profile, which could lead to a favorable effect during long-term treatment.

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