Orientador: Paulo Eduardo de Oliveira Carvalho / Coorientador: Antonio José Maria Catâneo / Banca: Donaldo Cerci da Cunha / Banca: Olavo Ribeiro Rodrigues / Banca: Marcos Vinícius Muriano da Silva / Banca: Gilberto Uemura / Resumo: Introdução: O câncer de mama metastático apresenta sobrevida inferior a 25% em cinco anos. A superexpressão do receptor HER2 nas células tumorais destas pacientes piora o prognóstico e diminui ainda mais sobrevida destas pacientes. O lapatinibe é uma droga que bloqueia a porção intracelular do receptor HER2 e esta ação poderia diminuir a proliferação das células neoplásicas diminuindo a progressão do tumor. Foi realizada uma revisão sistemática e metanálise dos estudos que avaliaram a eficácia e segurança do lapatinibe associado à quimioterapia ou terapia hormonal em mulheres com câncer de mama avançado com superexpressõ do HER2. Métodos: A busca dos estudos foi realizada nas bases MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. Foram analisados, como desfecho primário, a sobrevida global (SG), a sobrevida livre de progressão (SLP) e a qualidade de vida, e como desfechos secundários os efeitos colaterais relacionados ao tratamento. Resultados: Foram identificados e avaliados 1298 estudos. No final, aplicando os critérios de inclusão, foram selecionados cinco estudos. Juntos, estes estudos totalizaram 1127 pacientes com câncer de mama avançado (TNM estádio III ou IV) com superexpressão de HER2. Em ralação à SG foi observado que Lapatinibe associado a quimioterapia ou terapia hormonal diminui a probabilidade de morte em 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94;) p=0,005. Em relação à SLP o Lapatinibe diminui a probabilidade de eventos de progressão tumoral ou morte por qualquer causa em 48% (Hazard Ratio 0.58, 95% CI 0.50 to 0.67 p<0,0001. Nenhum estudo analisou a qualidade de vida. Referente aos efeitos colaterais, os pacientes que receberam Lapatinibe tiveram mais diarreia (RR 2,5, 95 IC 1,82 a 3,43; I2 =86 p <0,00001, aumento de risco de eventos cardíacos (RR 1,74, 95 % IC 1,02 a 2,99; I2 =0% p 0,04), e maior risco de sofrer descontinuidade de... / Abstract: Introduction: The five years survival of women with metastatic breast cancer accounts less than 25%. HER2 is an epidermal growth factor; the overexpression of HER2 in the cancer cells decreases survival of these patients Lapatinib is a drug that blocks the intracellular domain of HER2. This blocking could stop or diminish the cell proliferation and improve the survival of these patients. WE performed systematic review and metaanalysis of studies that analyzed the efficacy and safety of lapatinib associated with chemotherapy or hormone therapy for patients with advanced breast cancer that overexpress HER2. Methods: A search of the studies was conducted in MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. There were analyzed as the primary endpoint, overall survival (OS), progression-free survival (PFS) and the quality of life, and as secondary endpoints side effects, Results: We identified and assessed 1298 studies. Appling the inclusion criteria were selected five studies. Together these studies included a total of 1127 patients with advanced breast cancer (TNM stage III or IV) with overexpression of HER2. In conclusion it was observed that: for OS lapatinib combined with other drugs decreases the probability of death in 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94) p = 0.005. For PFS, lapatinib decreases the probability of tumor progression events or death from any cause in 48% (hazard ratio 0:58, 95% CI 0.67 to 0:50 P < 0,0001. None of the studies analyzed the quality of life. Refers to side effects, patients receiving Lapatinib had more diarrhea (RR 2.5, 95 CI 1.82 to 3.43; P<0.00001; increased risk of cardiac events (RR 1.74, 95% CI 1.02 the 2.99; P<0.04), and higher discontinuation of treatment (RR 4.00, 95% CI 1.44 the 11.17). Conclusion: The use of Lapatinib associated with other drugs decreases the probability of death and increases the likelihood of disease progression free ... / Doutor

Tumores de mama podem ser categorizados pela expressão de diferentes receptores, como por exemplo o HER-2 (receptor de fator de crescimento epidérmico 2), sendo amplamente estudada a utilização do fármaco inibidor do receptor da tirosina–quinase, ditosilato de lapatinib. Nesse sentido, o trabalho está centrado no desenvolvimento farmacotécnico e caracterização físico-química de nanocápsulas multiparede revestidas com polissorbato 80 e fosfatidilcolina (LIPOID S75®), seguidas de revestimento com quitosana, contendo ditosilato de lapatinib (25μg/mL e 50μg/mL). Estudos físico-químicos da fase orgânica e das nanocápsulas contendo LIPOID S75®, em diferentes concentrações, foram desenvolvidos através de medidas de viscosidade, tensão superficial e analise de calorimetria exploratória diferencial, correlacionando com o tamanho de partícula e interação da fosfatidilcolina no sistema. A partir disso, escolheu-se a maior concentração de LIPOID S75® testada, para desenvolver as nanocápsulas multiparede complexadas com metal e fenilalanina contendo ditosilato de lapatinib (Lap25-P5-Q-Fe-Phe; Lap50-P5-Q-Fe-Phe; Lap25-P5-Q-Zn-Phe; Lap50-P5-Q-Zn-Phe). As formulações contendo fármaco apresentaram distribuição unimodal de partículas na faixa nanométrica, (111 - 172 nm) e baixo índice de polidispersão. A faixa de potencial zeta obtida (+16 - +22 mV) indicou o revestimento da partícula pela quitosana. O ditosilato de lapatinib foi quantificado por cromatografia líquida de alta eficiência (HPLC). A fase móvel do método consistiu em acetonitrila:acetato de amônio 0,02M (70:30), vazão de 1,0 mL/min com detecção em 260 nm. As suspensões contendo 25 μg/mL e 50 μg/mL de fármaco apresentaram teor entre 93% e 91%, respectivamente. A eficiência de encapsulação (EE%) foi acima de 90%. O conjunto destes estudos demonstra que nanocápsulas desenvolvidas são potenciais candidatas para ensaios biológicos in vitro. / Breast tumors may be categorized by the expression of different receptors such as HER-2 (growth factor epidermal receptor 2), The use of the inhibitor drug receptor tyrosine-kinase lapatinib ditosylate (Tyverb®, GlaxoSmithKline) has been widely studied. In this sense, the work is focused on pharmaceutics development and physicochemical characterization of multiwall nanocapsules coated with polysorbate 80 and phosphatidylcholine (Lipoid S75®), followed by coating with chitosan containing lapatinib ditosylate (25 μ/mL and 50 μg/mL). Physicochemical studies of the organic phase and nanocapsules containing Lipoid S75® at different concentrations were developed by viscosity, surface tension analysis and differential scanning calorimetry, correlated with the particle size and interaction of phosphatidylcholine in the system From this, we have chosen the highest concentration of lipoid S75® tested to develop complexed nanocapsules multiparede with metal and phenylalanine containing lapatinib ditosylate (Lap25-P5-Phe-Phe-Q; Lap50-P5-Q -Fc-Phe; Lap25-P5-O-Zn-Phe; Lap50-P5-Zn-O-Phe). Formulations containing lapatinib ditosylate showed unimodal distribution of particles in the nanometer range (111-172 nm) and low polydispersity index. The obtained zeta potential range (+16 - +22 mV) indicated by the chitosan particle coating. The lapatinib ditosylate was quantified by high-performance liquid chromatography (HPLC). The mobile phase consisted method in acetonitrile: 0.02M ammonium acetate (70:30), flow 1.0 mLmin-1 with detection at 260 nm. The suspensions containing 25 ugmL-1 and 50 ugmL-1 showed drug content of between 93% and 91%, respectively. The encapsulation efficiency (EE%) was above 90%. Together these studies demonstrate that the nanocapsules are potential candidates for in vitro biological assays.

Made available in DSpace on 2016-08-12T18:48:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-11-26. Added 1 bitstream(s) on 2016-08-12T18:51:08Z : No. of bitstreams: 1 000864495.pdf: 561808 bytes, checksum: dff5ee77e1a3a3047af80805ca02be32 (MD5) / Introdução: O câncer de mama metastático apresenta sobrevida inferior a 25% em cinco anos. A superexpressão do receptor HER2 nas células tumorais destas pacientes piora o prognóstico e diminui ainda mais sobrevida destas pacientes. O lapatinibe é uma droga que bloqueia a porção intracelular do receptor HER2 e esta ação poderia diminuir a proliferação das células neoplásicas diminuindo a progressão do tumor. Foi realizada uma revisão sistemática e metanálise dos estudos que avaliaram a eficácia e segurança do lapatinibe associado à quimioterapia ou terapia hormonal em mulheres com câncer de mama avançado com superexpressõ do HER2. Métodos: A busca dos estudos foi realizada nas bases MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. Foram analisados, como desfecho primário, a sobrevida global (SG), a sobrevida livre de progressão (SLP) e a qualidade de vida, e como desfechos secundários os efeitos colaterais relacionados ao tratamento. Resultados: Foram identificados e avaliados 1298 estudos. No final, aplicando os critérios de inclusão, foram selecionados cinco estudos. Juntos, estes estudos totalizaram 1127 pacientes com câncer de mama avançado (TNM estádio III ou IV) com superexpressão de HER2. Em ralação à SG foi observado que Lapatinibe associado a quimioterapia ou terapia hormonal diminui a probabilidade de morte em 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94;) p=0,005. Em relação à SLP o Lapatinibe diminui a probabilidade de eventos de progressão tumoral ou morte por qualquer causa em 48% (Hazard Ratio 0.58, 95% CI 0.50 to 0.67 p<0,0001. Nenhum estudo analisou a qualidade de vida. Referente aos efeitos colaterais, os pacientes que receberam Lapatinibe tiveram mais diarreia (RR 2,5, 95 IC 1,82 a 3,43; I2 =86 p <0,00001, aumento de risco de eventos cardíacos (RR 1,74, 95 % IC 1,02 a 2,99; I2 =0% p 0,04), e maior risco de sofrer descontinuidade de... / Introduction: The five years survival of women with metastatic breast cancer accounts less than 25%. HER2 is an epidermal growth factor; the overexpression of HER2 in the cancer cells decreases survival of these patients Lapatinib is a drug that blocks the intracellular domain of HER2. This blocking could stop or diminish the cell proliferation and improve the survival of these patients. WE performed systematic review and metaanalysis of studies that analyzed the efficacy and safety of lapatinib associated with chemotherapy or hormone therapy for patients with advanced breast cancer that overexpress HER2. Methods: A search of the studies was conducted in MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. There were analyzed as the primary endpoint, overall survival (OS), progression-free survival (PFS) and the quality of life, and as secondary endpoints side effects, Results: We identified and assessed 1298 studies. Appling the inclusion criteria were selected five studies. Together these studies included a total of 1127 patients with advanced breast cancer (TNM stage III or IV) with overexpression of HER2. In conclusion it was observed that: for OS lapatinib combined with other drugs decreases the probability of death in 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94) p = 0.005. For PFS, lapatinib decreases the probability of tumor progression events or death from any cause in 48% (hazard ratio 0:58, 95% CI 0.67 to 0:50 P < 0,0001. None of the studies analyzed the quality of life. Refers to side effects, patients receiving Lapatinib had more diarrhea (RR 2.5, 95 CI 1.82 to 3.43; P<0.00001; increased risk of cardiac events (RR 1.74, 95% CI 1.02 the 2.99; P<0.04), and higher discontinuation of treatment (RR 4.00, 95% CI 1.44 the 11.17). Conclusion: The use of Lapatinib associated with other drugs decreases the probability of death and increases the likelihood of disease progression free ...

Tumores de mama podem ser categorizados pela expressão de diferentes receptores, como por exemplo o HER-2 (receptor de fator de crescimento epidérmico 2), sendo amplamente estudada a utilização do fármaco inibidor do receptor da tirosina–quinase, ditosilato de lapatinib. Nesse sentido, o trabalho está centrado no desenvolvimento farmacotécnico e caracterização físico-química de nanocápsulas multiparede revestidas com polissorbato 80 e fosfatidilcolina (LIPOID S75®), seguidas de revestimento com quitosana, contendo ditosilato de lapatinib (25μg/mL e 50μg/mL). Estudos físico-químicos da fase orgânica e das nanocápsulas contendo LIPOID S75®, em diferentes concentrações, foram desenvolvidos através de medidas de viscosidade, tensão superficial e analise de calorimetria exploratória diferencial, correlacionando com o tamanho de partícula e interação da fosfatidilcolina no sistema. A partir disso, escolheu-se a maior concentração de LIPOID S75® testada, para desenvolver as nanocápsulas multiparede complexadas com metal e fenilalanina contendo ditosilato de lapatinib (Lap25-P5-Q-Fe-Phe; Lap50-P5-Q-Fe-Phe; Lap25-P5-Q-Zn-Phe; Lap50-P5-Q-Zn-Phe). As formulações contendo fármaco apresentaram distribuição unimodal de partículas na faixa nanométrica, (111 - 172 nm) e baixo índice de polidispersão. A faixa de potencial zeta obtida (+16 - +22 mV) indicou o revestimento da partícula pela quitosana. O ditosilato de lapatinib foi quantificado por cromatografia líquida de alta eficiência (HPLC). A fase móvel do método consistiu em acetonitrila:acetato de amônio 0,02M (70:30), vazão de 1,0 mL/min com detecção em 260 nm. As suspensões contendo 25 μg/mL e 50 μg/mL de fármaco apresentaram teor entre 93% e 91%, respectivamente. A eficiência de encapsulação (EE%) foi acima de 90%. O conjunto destes estudos demonstra que nanocápsulas desenvolvidas são potenciais candidatas para ensaios biológicos in vitro. / Breast tumors may be categorized by the expression of different receptors such as HER-2 (growth factor epidermal receptor 2), The use of the inhibitor drug receptor tyrosine-kinase lapatinib ditosylate (Tyverb®, GlaxoSmithKline) has been widely studied. In this sense, the work is focused on pharmaceutics development and physicochemical characterization of multiwall nanocapsules coated with polysorbate 80 and phosphatidylcholine (Lipoid S75®), followed by coating with chitosan containing lapatinib ditosylate (25 μ/mL and 50 μg/mL). Physicochemical studies of the organic phase and nanocapsules containing Lipoid S75® at different concentrations were developed by viscosity, surface tension analysis and differential scanning calorimetry, correlated with the particle size and interaction of phosphatidylcholine in the system From this, we have chosen the highest concentration of lipoid S75® tested to develop complexed nanocapsules multiparede with metal and phenylalanine containing lapatinib ditosylate (Lap25-P5-Phe-Phe-Q; Lap50-P5-Q -Fc-Phe; Lap25-P5-O-Zn-Phe; Lap50-P5-Zn-O-Phe). Formulations containing lapatinib ditosylate showed unimodal distribution of particles in the nanometer range (111-172 nm) and low polydispersity index. The obtained zeta potential range (+16 - +22 mV) indicated by the chitosan particle coating. The lapatinib ditosylate was quantified by high-performance liquid chromatography (HPLC). The mobile phase consisted method in acetonitrile: 0.02M ammonium acetate (70:30), flow 1.0 mLmin-1 with detection at 260 nm. The suspensions containing 25 ugmL-1 and 50 ugmL-1 showed drug content of between 93% and 91%, respectively. The encapsulation efficiency (EE%) was above 90%. Together these studies demonstrate that the nanocapsules are potential candidates for in vitro biological assays.

La P-glycoprotéine (P-gp) est une protéine transmembranaire de la famille des ATP binding cassette transporteurs. Elle est impliquée dans l'efflux du milieu intracellulaire vers le milieu extracellulaire d'une grande variété de médicaments anticancéreux. Elle peut être responsable de la diminution de la biodisponibilité orale et de la concentration intra-tumorale des médicaments qui en sont substrats. Elle peut notamment être surexprimée par les cellules cancéreuses des adénocarcinomes du colon naïfs de tout traitement, suggérant une résistance naturelle de cette tumeur et également après une chimiothérapie. Notre premier travail in vivo a documenté le caractère substrat de la P-gp de l'evérolimus, inhibiteur de mTOR indiqué dans divers cancers (rein, tumeurs neuroendocrines d'oringine pancréatique et sein), jusqu'à maintenant uniquement étudié dans des modèles in vitro. Une augmentation significative de l'AUC de l'evérolimus administré par voie orale est observée chez des souris mdr1a-/b- comparées à des souris mdr1a+/1b+. Une amélioration significative de la biodisponibilité orale de l'evérolimus est aussi notée chez des souris prétraitées par le lapatinib (Tyverb®), inhibiteur des tyrosines kinases (EGFR et HER2) indiqué dans le cancer du sein, par rapport aux souris ayant reçu l'evérolimus seul. Ce résultat est accompagné d'une inhibition de l'expression de la P-gp intestinale par le lapatinib mesurée par la technique de Western Blot. Enfin, une étude préclinique menée chez des souris porteuses d'une xénogreffe colorectale mutée KRAS montre une activité anti-tumorale certaine des deux médicaments utilisés seuls et en schéma séquentiel. Notre seconde étude a montré pour la première fois que le cétuximab (Erbitux®), anticorps anti-EGFR, inhibe la fonctionnalité de la P-gp dans deux lignées cellulaires surexprimant la P-gp (les cellules IGROV-1 et les HEK P-gp) indépendamment de leur statut EGFR et entraîne chez des souris porteuses d'une xénogreffe colorectale une augmentation significative de la biodisponibilité orale et de la concentration intra-tumorale du SN-38, métabolite actif de l'irinotécan (Campto®) administré par voie orale. Le cétuximab étant prescrit en association avec l'irinotécan chez des patients atteints d'un cancer colorectal métastasé, initialement réfractaire à l'irinotécan, ces résultats pourraient en partie expliquer la réversion de la résistance à l'irinotécan par le cétuximab par une inhibition de l'efflux de la P-gp. Grâce à l'étude de deux associations de médicaments "lapatinib-evérolimus" et "cétuximab-irinotécan", nous avons démontré l'intérêt de l'étude de l'inhibition de la P-gp avec les traitements les plus récents, notamment son rôle dans l'amélioration de la biodisponibilité orale de chimiothérapies utilisées par voie orale.

Carvacrol is a phenol monoterpene found in aromatic plants specially in Lamiaceae family, which has been evaluated in an experimental model of breast cancer. However, any proposed mechanism based on its antitumor effect has not been reported. In our previous study, carvacrol showed a protective effect on 7,12-dimethylbenz[α]anthracene- (DMBA-) induced breast cancer in female rats. The main objective in this research was to evaluate by using in silico study the carvacrol on HER2, PI3Kα, mTOR, hERα, PR, and EGFR receptors involved in breast cancer progression by docking analysis, molecular dynamic, and drug-likeness evaluation. A multilevel computational study to evaluate the antitumor potential of carvacrol focusing on the main targets involved in the breast cancer was carried out. The in silico study starts with protein-ligand docking of carvacrol followed by ligand pathway calculations, molecular dynamic simulations, and molecular mechanics energies combined with the Poisson–Boltzmann (MM/PBSA) calculation of the free energy of binding for carvacrol. As result, the in silico study led to the identification of carvacrol with strong binding affinity on mTOR receptor. Additionally, in silico drug-likeness index for carvacrol showed a good predicted therapeutic profile of druggability. Our findings suggest that mTOR signaling pathway could be responsible for its preventive effect in the breast cancer. / Revisión por pares

Το γλοιοβλάστωμα αποτελεί έναν από τους πιο θανατηφόρους τύπους καρκίνου για τον άνθρωπο δεδομένου ότι ο μέσος όρος επιβίωσης είναι 12-15 μήνες. Η συμβατική θεραπεία με τα κλασσικά χημειοθεραπευτικά σχήματα δεν έχει αποδώσει ιδιαιτέρως θετικά αποτελέσματα. Η εξέλιξη της μοριακής βιολογίας έχει «ρίξει φως» στην διερεύνηση και κατανόηση αρκετών μηχανισμών της κυτταρικής λειτουργίας που αφορούν τη μετανάστευση, τον πολλαπλασιασμό και την απόπτωση των κυττάρων. Στο γλοιοβλάστωμα έχει παρατηρηθεί αυξημένη δραστηριότητα πολλών υποδοχέων αυξητικών παραγόντων στην επιφάνεια των κυττάρων όπως ο EGFR, ο PDGFR και ο VEGFR. Ταυτόχρονα, υπάρχουν και άλλα μόρια επιφανείας με ιδιαίτερο ενδιαφέρον όπως οι ιντεγκρίνες λόγω της ιδιότητας τους να καθορίζουν τη μετανάστευση. Στη νέα εποχή των αναστολέων κινάσης τυροσίνης οι οποίοι δρουν έναντι των υποδοχέων των αυξητικών παραγόντων και θεωρούνται πολλά υποσχόμενα μικρά μόρια, πρέπει να διερευνηθεί αν μπορούν να έχουν θέση στην αντιμετώπιση του γλοιοβλαστώματος. Αρχικά έγινε εκτίμηση της επίδρασης των αναστολέων sunitinib και lapatinib ξεχωριστά αλλά και σε συνδυασμό, στο πολλαπλασιασμό και στο κυτταρικό θάνατο των κυττάρων γλοιβλαστώματος U87 και M059K σε συγκεντρώσεις 0,01, 0,1, 1 και 10μΜ. Για το σκοπό αυτό χρησιμοποιήθηκε η μέθοδος του 3-[4,5-dimethylthiazol-2-yl]-2,5 διμεθυλθειαζόλο βρωμιδίου και το ολοκληρωμένο σύστημα ανίχνευσης αννεξίνης V/προπιδίου (annexin V/propidium iodide), αντίστοιχα. Για τη διαδικασία της μετανάστευσης εφαρμόστηκε η μέθοδος μελέτης του χημειοτακτισμού. Η έκφραση μεταλλοπρωτεϊνασών MM-2 και MMP-9 εκτιμήθηκε με τη μέθοδο του ζυμογραφήματος. Περαιτέρω έγινε διερεύνηση της πιθανής εμπλοκής των φαρμάκων στο σχηματισμό συμπλόκων EGFR-υπομονάδα β1 ιντεγκρινών, PDGFR- υπομονάδα β3 ιντεγκρινών και VEGFR- υπομονάδα β3 ιντεγκρινών. Η μελέτη της δημιουργίας των συμπλόκων των β υπομονάδων των ιντεγκρινών με τους υποδοχείς αυξητικών παραγόντων έγινε χρησιμοποιώντας τη μέθοδο της ανοσοκατακρήμνισης και της ανάλυσης κατά Western. Τα αποτελέσματα επαληθεύτηκαν και με τη μέθοδο του ανοσοφθορισμού. Με την ίδια μέθοδο μελετήθηκε και η επίδραση των φαρμάκων στη φωσφορυλιωμένη μορφή της FAK. Τα πειραματικά δεδομένα έδειξαν ότι και οι δύο αναστολείς sunitinib και lapatinib, μείωσαν τον πολλαπλασιασμό με δοσοεξαρτώμενο τρόπο 48 ώρες μετά την προσθήκη τους στα κύτταρα είτε μεμονωμένα είτε σε συνδυασμό. Τα αποτελέσματα της αναστολής του πολλαπλασιασμού συμβάδιζαν με τα αποτελέσματα της απόπτωσης όπου το ποσοστό των αποπτωτικών κυττάρων αυξήθηκε. Περαιτέρω η μετανάστευση των κυττάρων εμφανίστηκε μειωμένη μετά την προσθήκη των φαρμάκων είτε μεμονωμένα είτε σε συνδυασμό. Η έκφραση των μεταλλοπρωτεϊνασών MMP-2 και MMP-9 δεν επηρεάστηκε στα κύτταρα U87 μετά την προσθήκη και των 2 φαρμάκων. Αντίθετα, στα κύτταρα M059K το sunitinib αλλά και ο συνδυασμός του με το lapatinib ελαττώσαν την έκφραση των MMPs 48 ώρες μετά τη προσθήκη τους στο θρεπτικό μέσο. Στη συνέχεια βρέθηκε ότι το lapatinib μπορεί να αναστείλει την δημιουργία συμπλόκου του EGFR με την υπομονάδα β1 των ιντεγκρινών, έως και 30 λεπτά μετά την προσθήκη του φαρμάκου στα κύτταρα. Αντίστοιχα το sunitinib μπορεί να αναστέλλει το σχηματισμό συμπλόκου της υπομονάδας β3 των ιντεγκρινών με τον VEGFR εντός δύο ωρών από την προσθήκη των φαρμάκων, χωρίς επίδραση όμως στον σχηματισμό των συμπλόκων PDGFR – υπομονάδα β3 ιντεγκρίνης. Αυτά τα αποτελέσματα επιβεβαιώθηκαν και με την χρήση ανοσοφθορισμού. Συνοψίζοντας, οι δύο αναστολείς κινάσης τυροσίνης, sunitinib και lapatinib κατάφεραν να επιδείξουν αξιόλογα αποτελέσματα στις παραμέτρους του πολλαπλασιασμού και της μετανάστευσης των κυττάρων γλοιοβλαστώματος. Τα αποτελέσματα της έρευνας είναι πρωτοποριακά διότι, αναφορικά με την μετανάστευση στο γλοίωμα, αυτή φαίνεται να αναστέλλεται αποτελεσματικότερα με τον συνδυασμό των δύο φαρμάκων πιθανά μέσω του μηχανισμού διαταραχής της δημιουργίας του συμπλόκου ιντεγκρίνης- υποδοχέας αυξητικού παράγοντα. / Glioblastoma is considered to be one of the most fatal among the malignancies in human with median survival between 12-15 months. The conventional chemotherapy cannot change the fate of these patients. Recent advances in molecular biology have shed light in the various mechanisms recruited by malignant cells in order to proliferate, metastasize and escape apoptosis. Studies in glioblastoma have already shown up-regulation in the function of tyrosine kinase receptors such as EGFR, PDGFR and VEGFR. Besides this, in the more recent years, research supports the significant role of integrins in the migration process of glioblastoma cells. In the new era of tyrosine kinase inhibitors (TKIs), where their main mechanism of action is by blocking various growth factor receptors, it has to be determined a possible contribution in the fight against glioblastoma. Initially, it was estimated a possible effect of sunitinib and lapatinib applied either alone or in combination, on U87 and M059K glioma cells’ proliferation and apoptosis using doses of 0,01 μM, 0,1 μM, 1μM and 10 μM. The proliferation was estimated using the 3-[4,5-dimethylthiazol-2-yl]-2,5 dimethyltetrazolium bromide assay and apoptosis using annexin V/propidium iodide detection assay. Migration assay was performed using Boyden chamber assay. The release of MMP-2 and MMP-9 into the culture medium of U87 and M059K cells was measured by zymography. Furthermore, a possible implication of the tested agents in the formation of EGFR-integrin β1 complex, VEGFR-integrin β3 and PDGFR-integrin β3 complexes formation was studied. Immunoprecipitation and western blot analysis were used for studying the complex formation of EGFR, PDGFR and VEGFR with integrins. Validation of the results was made using immunofluorescence assay. Also, similar experiments were performed for the effect of sunitinib and lapatinib in FAK phosphorylation. The results showed that both tested agents decreased cell proliferation in a dose-dependent manner 48 h after their application in both cell lines either alone or in combination. The results in cell proliferation were in line with the increase in apoptotic cells after their treatment with the tested agents. Furthermore, the ability of U87 and M059K cells to migrate was inhibited either by each agent alone or in combination. Both agents did not affect MMP-2 and MMP-9 levels in U87 cells, however, MMPs levels were decreased in M059K cells, 48h after their treatment with sunitinib either alone or in combination with lapatinib. Additionally, a time course study for the effect of lapatinib on EGFR-integrin β1 complex revealed an inhibition in complex formation up to 30 min after agent application. Likewise, sunitinib inhibited complex formation of VEGFR-integrin β3 complex within 2h after its application without affecting PDGFR-integrin β3 complex. The previously-described interruption of complexes formation was confirmed with an immunofluorescence assay. Summarizing the results, sunitinib and lapatinib exerted significant effects on glioblastoma cell proliferation, apoptosis and migration. The preliminary results of the current study are the first to support the implication of a dual anti-EGFR/HER-2 agent, lapatinib and a multi-targeted agent, sunitinib in glioma cells migration, through a mechanism implying interruption of growth factor receptor - integrin complexes formation.