Background LHRH agonists decrease tumour size/activity by suppressing testosterone in prostate cancer; however, initial injection causes testosterone surge that triggers flare symptoms. Anti-androgen given with agonist may reduce/avoid flare symptoms. When LHRH antagonist/blocker is introduced, testosterone suppression is immediate, but there is uncertainty about significance of flare symptoms without anti-androgen. Objective Systematic review compared significance of flare symptoms avoided and cost utility analysis using modelling comparing incremental value of blocker (degarelix) OR agonist (goserelin)+anti-androgen (bicalutamide) VERSUS agonist alone in prostate cancer patients. Outcome Incremental cost/QALY of bone pain as flare symptom between treatments Results Thirteen studies were reviewed. There was no standard definition for flare symptoms or data on LHRH antagonist versus other treatments on flare. From societal perspective, goserelin+bicalutamide was dominated over goserelin alone and similarly, from public perspective, goserelin+bicalutamide had favourable cost effectiveness profile against goserelin. Conclusion With bone pain as clinical endpoint, LHRH agonist+anti-androgen had favourable cost-effectiveness profile compared to goserelin.

Background LHRH agonists decrease tumour size/activity by suppressing testosterone in prostate cancer; however, initial injection causes testosterone surge that triggers flare symptoms. Anti-androgen given with agonist may reduce/avoid flare symptoms. When LHRH antagonist/blocker is introduced, testosterone suppression is immediate, but there is uncertainty about significance of flare symptoms without anti-androgen. Objective Systematic review compared significance of flare symptoms avoided and cost utility analysis using modelling comparing incremental value of blocker (degarelix) OR agonist (goserelin)+anti-androgen (bicalutamide) VERSUS agonist alone in prostate cancer patients. Outcome Incremental cost/QALY of bone pain as flare symptom between treatments Results Thirteen studies were reviewed. There was no standard definition for flare symptoms or data on LHRH antagonist versus other treatments on flare. From societal perspective, goserelin+bicalutamide was dominated over goserelin alone and similarly, from public perspective, goserelin+bicalutamide had favourable cost effectiveness profile against goserelin. Conclusion With bone pain as clinical endpoint, LHRH agonist+anti-androgen had favourable cost-effectiveness profile compared to goserelin.

The decapeptide, gonadotropin releasing hormone (GnRH), is the central regulator of reproductive function. It binds to receptors on the gonadotrope cells of the pituitary and stimulates release of luteinizing hormone (LH) and follicle stimulating hormone (FSH). Eleven different structural forms of GnRH have now been identified in various animal species. Chimaeric analogues of some of the variant forms of GnRH were synthesized in order to study the functional significance of the most common amino acid substitutions, which occur in positions 5, 7 and 8. Peptide binding affinities for sheep and rat GnRH receptors and potencies in stimulating LH and FSH release from cultured sheep pituitary cells and LH release from cultured chicken pituitary cells were measured. Histidine in position 5 decreased LH releasing potency in chicken cells, but slightly increased receptor binding affinity in rat and sheep membranes. Tryptophan in position 7 had minimal effect on GnRH activity in mammals, but increased LH release in chicken cells. Although differences in the structural requirements of mammalian and chicken GnRH receptors were anticipated, it was also found that rat GnRH receptors exhibited higher affinity for analogues with Tryptophan in position 7, than did sheep GnRH receptors. Substitutions in position 8 revealed the most marked differences in the structural requirements of mammalian and chicken GnRH receptors. Arginine was required for high GnRH activity in mammalian systems, but analogues with neutral substitutions in position 8 were more potent in chicken pituitary cells. The tolerance of position 8 substitutions, combined with the relatively small effects, in chicken cells, of incorporating a D-amino acid in position 6, indicate that the chicken GnRH receptor is less stringent than mammalian receptors in its recognition of peptide conformation. To examine how changes in ligand structure cause changes in receptor binding affinity and receptor activation, it was necessary to know the structures of the GnRH receptors. A protocol was developed for the purification of GnRH binding proteins from detergent-solubilized pituitary membranes, by affinity chromatography. This procedure yielded a protein which migrated as a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis, but was different from the recently cloned GnRH receptor. To test the proposal that the arginine residue in mammalian GnRH interacts with an acidic receptor residue, eight conserved acidic residues of the cloned mouse GnRH receptor were mutated to asparagine or glutamine. Mutant receptors were transiently expressed in COS-1 cells and tested for decreased preference for Arg⁸-containing ligands by ligand binding and inositol phosphate production. One mutant receptor, in which the glutamate residue in position 301 was mutated, exhibited decreased affinity for mammalian GnRH. The mutant receptor also exhibited decreased affinity for [Lys⁸]-GnRH, but unchanged affinity for [Gln⁸]-GnRH compared with the wildtype receptor, and increased affinity for the acidic analogue, [Glu⁸]-GnRH. This loss of affinity was specific for the residue in position 8, because the mutant receptor retained hiszh affinity for analogues with favourable substitutions in positions 5, 6 and 7. Thus, the Glu³⁰¹ residue of the GnRH receptor plays a role in receptor recognition of Arg⁸ in the ligand, consistent with an electrostatic interaction between these two residues. The Glu³⁰¹ and Arg⁸ residues were not required for the high affinity interactions of conformationally constrained peptides. This indicates that an interaction which involves these two residues may induce changes in the conformation of GnRH after it has bound to the receptor.

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.

The existence of an ultrashort feedback mechanism regulating GnRH secretion has been supported from in vivo and in vitro studies. However, the complex synaptic connections of GnRH neurons with other neural elements made it difficult to determine whether the regulation was mediated by direct actions on the GnRH neurons or through actions on other interneurons. The recent development of the GnRH-secreting neuronal cell line, GT1, provided a model system for the study of neural regulation of a pure population of GnRH neurons. The present studies utilized GT1 -7 cells to investigate whether GnRH (at the level of the nerve terminal) influences the control of its own release. Preliminary studies determined the presence of GnRH mRNA in GT1-7 cells and established a cell culture system for the analysis of secretagogue-induced GnRH release. In this system GnRH release was shown to be spontaneous and was enhanced by the addition of K⁺, L-GLU, forskolin and PMA. Furthermore, K⁺- and forskolin-induced GnRH release was dependent on extracellular Ca²⁺. For the analysis of an ultrashort feedback mechanism, GT1-7 cells were cultured in 6-well plates to near confluence and then incubated in serum-free medium in the presence (1 nM- 1 μM) or absence of GnRH antagonist, Ant 27. Basal, K⁺-and forskolin-induced secretion of GnRH was monitored with antiserum 1076 which does not cross-react with Ant 27 at> 1 μM. Ant 27 treatment increased basal, K⁺- and forskolin-stimulated GnRH release in a dose-dependent manner. Total content was unaffected by 18 h treatment of GT1-7 cells with Ant 27. This suggests that the effects of Ant 27 are at the level of release and not biosynthesis. The presence of GnRH binding sites in the cells was demonstrated with ¹²⁵I-GnRH analog. These findings support the concept that GnRH, acting via autoreceptors, negatively controls its own release.

In the present study our objectives were (1) to follow the temporal patterns of testicular LH and FSH receptor (LH-R and FSH-R) concentrations and affinity (Ka) during sexual maturation in bulls, to see if such patterns could explain the control of rapid testicular growth that occurs after 25 weeks of age, when serum gonadotropin concentrations are low; (2) to see if transformation growth factors (TGF- alpha and beta 1, 2 and 3) and interleukins (IL-1 and IL-6) are produced in the developing bovine testis and if their concentrations change during development; (3) to see if the onset of puberty could be hastened by treating bull calves subcutaneously (sc) with 3 mg of bLH (n=6) or 4 mg of bFSH (n=6) once every 2 days, from 4 to 8 weeks after birth. Mean LH-R concentrations decreased from 13 to 25 weeks of age and increased to 56 weeks of age (P<0.05). LH-RKa decreased from 9 to 17 weeks of age, increased to 29 weeks and declined to 33 weeks of age (P<0.05). FSH-R concentrations declined from 17 to 25 weeks of age then increased to 56 weeks of age (P<0.05). FSH-RKa increased from 17 to 25 weeks of age (P<0.05). Testicular TGF-alpha concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks and from 33 to 56 weeks of age (P<0.05). Testicular TGF-beta 1 concentrations decreased from 17 to 21 weeks of age, increased to 25 weeks and decreased from 25 to 29 weeks of age (P<0.05). Testicular TGF-beta 2 concentrations increased from 5 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks and decreased at 29 weeks of age (P<0.05). Testicular TGF-beta 3 concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks of age and from 25 to 29 weeks of age (P<0.05). Mean testicular IL-1 alpha concentrations decreased from 5 to 9 weeks of age and 13 to 21 weeks of age (P<0.01) while mean testicular IL-1 beta concentrations decreased from 13 to 17 weeks and 29 to 33 weeks of age (P<0.01). Mean IL-1 bioactivity increased from 13 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks, decreased to 29 weeks and decreased from 33 to 56 weeks of age (P<0.05). Mean testicular IL-6 concentrations decreased (P<0.05) from 9 to 13 weeks of age, increased (P<0.05) to 21 weeks, decreased (P<0.05) to 25 weeks of age, increased (P<0.05) to 29 weeks and decreased (P<0.01) to 56 weeks of age. <p>We concluded that high concentrations of gonadotropin receptors might be critical to initiate postnatal testis growth and support it after 25 weeks of age in the face of low serum gonadotropin concentrations. Testicular TGF-alpha concentrations were higher in calves than adults while concentrations of TGF-beta and IL-1 were higher in the early postnatal period than the peripubertal period. The changes in testicular concentrations of TGFs and ILs led us to suggest a possible local regulatory role in development. Testicular IL-6 concentrations were higher in prepubertal calves than adults. Treatment of bull calves with bFSH from 4 to 8 weeks of age increased testicular growth (SC), hastened the onset of puberty (SC ≥ 28 cm), and enhanced spermatogenesis.

In the present study our objectives were (1) to follow the temporal patterns of testicular LH and FSH receptor (LH-R and FSH-R) concentrations and affinity (Ka) during sexual maturation in bulls, to see if such patterns could explain the control of rapid testicular growth that occurs after 25 weeks of age, when serum gonadotropin concentrations are low; (2) to see if transformation growth factors (TGF- alpha and beta 1, 2 and 3) and interleukins (IL-1 and IL-6) are produced in the developing bovine testis and if their concentrations change during development; (3) to see if the onset of puberty could be hastened by treating bull calves subcutaneously (sc) with 3 mg of bLH (n=6) or 4 mg of bFSH (n=6) once every 2 days, from 4 to 8 weeks after birth. Mean LH-R concentrations decreased from 13 to 25 weeks of age and increased to 56 weeks of age (P<0.05). LH-RKa decreased from 9 to 17 weeks of age, increased to 29 weeks and declined to 33 weeks of age (P<0.05). FSH-R concentrations declined from 17 to 25 weeks of age then increased to 56 weeks of age (P<0.05). FSH-RKa increased from 17 to 25 weeks of age (P<0.05). Testicular TGF-alpha concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks and from 33 to 56 weeks of age (P<0.05). Testicular TGF-beta 1 concentrations decreased from 17 to 21 weeks of age, increased to 25 weeks and decreased from 25 to 29 weeks of age (P<0.05). Testicular TGF-beta 2 concentrations increased from 5 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks and decreased at 29 weeks of age (P<0.05). Testicular TGF-beta 3 concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks of age and from 25 to 29 weeks of age (P<0.05). Mean testicular IL-1 alpha concentrations decreased from 5 to 9 weeks of age and 13 to 21 weeks of age (P<0.01) while mean testicular IL-1 beta concentrations decreased from 13 to 17 weeks and 29 to 33 weeks of age (P<0.01). Mean IL-1 bioactivity increased from 13 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks, decreased to 29 weeks and decreased from 33 to 56 weeks of age (P<0.05). Mean testicular IL-6 concentrations decreased (P<0.05) from 9 to 13 weeks of age, increased (P<0.05) to 21 weeks, decreased (P<0.05) to 25 weeks of age, increased (P<0.05) to 29 weeks and decreased (P<0.01) to 56 weeks of age. <p>We concluded that high concentrations of gonadotropin receptors might be critical to initiate postnatal testis growth and support it after 25 weeks of age in the face of low serum gonadotropin concentrations. Testicular TGF-alpha concentrations were higher in calves than adults while concentrations of TGF-beta and IL-1 were higher in the early postnatal period than the peripubertal period. The changes in testicular concentrations of TGFs and ILs led us to suggest a possible local regulatory role in development. Testicular IL-6 concentrations were higher in prepubertal calves than adults. Treatment of bull calves with bFSH from 4 to 8 weeks of age increased testicular growth (SC), hastened the onset of puberty (SC ≥ 28 cm), and enhanced spermatogenesis.

Thesis (doctoral)--Uppsala University, 1983. / Includes bibliographical references (p. 37-48).

No description available.

Die Überlebensrate von Patienten mit duktalem Adenokarzinom des Pankreas ist sowohl in primär resektablen als auch im lokal fortgeschrittenen und metastasierten Stadium kurz. Das duktale Adenokarzinom des Pankreas breitet sich rasch aus und wird aufgrund fehlender Frühsymptome oft erst im fortgeschrittenen Stadium diagnostiziert. Gegenwärtig fehlen spezifische Tumormarker, die eine frühzeitige Diagnose erlauben würden. Aufgrund zahlreicher Mechanismen der primären und sekundären Chemoresistenz ist das Pankreaskarzinom verhältnismäßig resistent gegenüber konventioneller systemisch verabreichter Chemotherapie, antikörperbasierten sowie niedermolekularen Therapiestrategien, Enzyminhibitoren, Bestrahlung und chirurgischer Therapie. Ein vielversprechender Angriffspunkt zur zielgerichteten Therapie des Pankreaskarzinoms eröffnet die tumorspezifische Expression des GnRH-I Rezeptors. In dieser Arbeit konnte der GnRH-IRezeptor durch RT-PCR und immunhistochemisch in 32,5% der duktalen Adenokarzinome des Pankreas nachgewiesen werden. Es wurde gezeigt, dass die Behandlung mit dem Hybridwirksoff AESZ-108 (AN-152), einem zytotoxischen GnRH-Analogon in vitro und in vivo Apoptose in den GnRH-IRezeptor- positiven Pankreaskarzinomzelllinien induziert. Apoptose wurde durch den intrinsischen Signalweg über den Zusammenbruch des mitochondrialen Membranpotentials vermittelt und führte zu DNA-Fragmentierung des Nukleus wie fluoreszenzmikroskopisch gezeigt werden konnte. Das zytotoxische GnRH-Analogon AESZ-108 (AN-152) führt in vivo zu einer signifikanten Inhibition des Tumorwachstums im Vergleich zur Therapie mit dem Anthrazyklin Doxorubicin, welches zu keiner signifikanten Inhibition des Pankreaskarzinomwachstums führt. Die rezeptorvermittelte Aufnahme ermöglicht eine selektive Therapie. Nach rezeptorvermittelter Endozytose wird das an die D-Lys6-Seitenkette gebundene Doxorubicin spezifisch im Nukleus der rezeptorpositiven Karzinomzellen freigesetzt. Die Ergebnisse dieser Arbeiten zeigen, dass AESZ-108 (AN-152) ein geeigneter Ansatz zur selektiven Chemotherapie GnRH-I-Rezeptor positiver humaner duktaler Adenokazinome des Pankreas ist.