RESUMO
The tumour microenvironment may contribute to tumorigenesis owing to mechanical forces such as fibrotic stiffness or mechanical pressure caused by the expansion of hyper-proliferative cells. Here we explore the contribution of the mechanical pressure exerted by tumour growth onto non-tumorous adjacent epithelium. In the early stage of mouse colon tumour development in the Notch(+)Apc(+/1638N) mouse model, we observed mechanistic pressure stress in the non-tumorous epithelial cells caused by hyper-proliferative adjacent crypts overexpressing active Notch, which is associated with increased Ret and ß-catenin signalling. We thus developed a method that allows the delivery of a defined mechanical pressure in vivo, by subcutaneously inserting a magnet close to the mouse colon. The implanted magnet generated a magnetic force on ultra-magnetic liposomes, stabilized in the mesenchymal cells of the connective tissue surrounding colonic crypts after intravenous injection. The magnetically induced pressure quantitatively mimicked the endogenous early tumour growth stress in the order of 1,200 Pa, without affecting tissue stiffness, as monitored by ultrasound strain imaging and shear wave elastography. The exertion of pressure mimicking that of tumour growth led to rapid Ret activation and downstream phosphorylation of ß-catenin on Tyr654, imparing its interaction with the E-cadherin in adherens junctions, and which was followed by ß-catenin nuclear translocation after 15 days. As a consequence, increased expression of ß-catenin-target genes was observed at 1 month, together with crypt enlargement accompanying the formation of early tumorous aberrant crypt foci. Mechanical activation of the tumorigenic ß-catenin pathway suggests unexplored modes of tumour propagation based on mechanical signalling pathways in healthy epithelial cells surrounding the tumour, which may contribute to tumour heterogeneity.
Assuntos
Carcinogênese/patologia , Neoplasias do Colo/fisiopatologia , Pressão , Microambiente Tumoral , beta Catenina/genética , Transporte Ativo do Núcleo Celular , Animais , Células Epiteliais/citologia , Células Epiteliais/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Imãs , Masculino , Nanopartículas Metálicas , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação , Proteínas Proto-Oncogênicas c-ret/metabolismo , Receptores Notch/genética , Receptores Notch/metabolismo , Transdução de Sinais , beta Catenina/metabolismoRESUMO
BACKGROUND: Colonic self-expanding metallic stents (SEMS) are used in obstructive colorectal cancer patients as a bridge to surgery. However, its oncologic safety remains uncertain. Therefore, we attempted to clarify this further with an experimental study and constructed a mouse model of colonic cancer. METHODS: CT26 cells were injected in the rectal wall, and to mimic SEMS, a cardiac stent was inserted under endoscopy in occlusive (75 % lumen occlusion) tumors. We set up a control group (n = 22) and a stent group (n = 16), and the findings were compared. We focused on serum lactate dehydrogenase (LDH) concentrations, circulating tumor cells, survival time, peritoneal carcinomatosis, liver metastases, and bioluminescence. RESULTS: One week after stent insertion, the serum LDH concentrations were significantly higher in the stent group (506 ± 203 IU/L) compared to the controls (229 ± 52 IU/L) (P = 0.005). The average survival time before sacrifice was significantly lower in the stent group (15.2 ± 1 days) compared to the controls (20 ± 5 days) (P = 0.005). The presence of a peritoneal carcinomatosis was more frequently observed in the stent group (75 %) than in the controls (50 %). Liver metastases were observed in 19 % of the stent group compared to the controls (4.5 %) (P = 0.29). After multivariate analysis, the stent group was still found to be associated with significantly lower survival time (P = 0.002). CONCLUSIONS: These observations led us to conclude that in our mouse model, SEMS resulted in an increased metastatic process and a shorter survival time. We suggest, therefore, that the utmost caution be exercised when opting for a stent as a bridge to surgery.
Assuntos
Neoplasias do Colo/cirurgia , Modelos Animais de Doenças , Obstrução Intestinal/cirurgia , Neoplasias Hepáticas/secundário , Neoplasias Peritoneais/secundário , Stents/efeitos adversos , Animais , Neoplasias do Colo/patologia , Humanos , CamundongosRESUMO
The main drawback of the anticancer chemotherapy consists in the lack of drug selectivity causing severe side effects. The targeted drug delivery appears to be a very promising strategy for controlling the biodistribution of the cytotoxic agent only on malignant tissues by linking it to tumor-targeting moiety. Here we exploit the natural characteristics of Shiga toxin B sub-unit (STxB) as targeting carrier on Gb3-positive cancer cells. Two cytotoxic conjugates STxB-doxorubicin (STxB-Doxo) and STxB-monomethyl auristatin F (STxB-MMAF) were synthesised using copper-free 'click' chemistry. Both conjugates were obtained in very high yield and demonstrated strong tumor inhibition activity in a nanomolar range on Gb3-positive cells.
Assuntos
Antineoplásicos/química , Química Click , Doxorrubicina/química , Portadores de Fármacos/química , Oligopeptídeos/química , Toxina Shiga/química , Anticorpos/imunologia , Antineoplásicos/síntese química , Antineoplásicos/toxicidade , Transporte Biológico , Sobrevivência Celular/efeitos dos fármacos , Cobre/química , Doxorrubicina/toxicidade , Portadores de Fármacos/síntese química , Desenho de Fármacos , Células HT29 , Células HeLa , Humanos , Microscopia Confocal , Oligopeptídeos/toxicidade , Toxina Shiga/imunologia , Toxina Shiga/metabolismoRESUMO
A key challenge in anticancer therapy is to gain control over the biodistribution of cytotoxic drugs. The most promising strategy consists in conjugating drugs to tumor-targeting carriers, thereby combining high cytotoxic activity and specific delivery. To target Gb3-positive cancer cells, we exploit the non-toxic B-subunit of Shiga toxin (STxB). Here, we have conjugated STxB to highly potent auristatin derivatives (MMA). A former linker was optimized to ensure proper drug-release upon reaching reducing environments in target cells, followed by a self-immolation step. Two conjugates were successfully obtained, and in vitro assays demonstrated the potential of this targeting system for the selective elimination of Gb3-positive tumors.
Assuntos
Aminobenzoatos/química , Antineoplásicos/química , Portadores de Fármacos/química , Oligopeptídeos/química , Toxina Shiga/química , Células HT29 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Conformação ProteicaRESUMO
The natural flavonoid fisetin (3,3',4',7-tetrahydroxyflavone) has shown antiangiogenic and anticancer properties. Because of fisetin limited water solubility, we designed a liposomal formulation and evaluated its biological properties in vitro and in Lewis lung carcinoma (LLC) bearing mice. A liposomal formulation was developed with DOPC and DODA-PEG2000, possessing a diameter in the nanometer range (173.5±2.4nm), a high homogeneity (polydispersity index 0.181±0.016) and high fisetin encapsulation (58%). Liposomal fisetin incubated with LLC cells were internalized, induced a typical fisetin morphological effect and increased the sub-G1 cell distribution. In vivo, liposomal fisetin allowed a 47-fold increase in relative bioavailability compared to free fisetin. The effect of liposomal fisetin on LLC tumor growth in mice at low dose (21mg/kg) allowed a higher tumor growth delay (3.3 days) compared to free fisetin at the same dose (1.6 day). Optimization of liposomal fisetin therapy was attempted by co-treatment with cyclophosphamide which led to a significant improvement in tumor growth delay (7.2 days) compared to cyclophosphamide with control liposomes (4.2 days). In conclusion, fisetin liposomes markedly improved fisetin bioavailability and anticancer efficacy in mice and this formulation could facilitate the administration of this flavonoid in the clinical setting.
Assuntos
Antineoplásicos/administração & dosagem , Antineoplásicos/farmacocinética , Carcinoma Pulmonar de Lewis/tratamento farmacológico , Flavonoides/administração & dosagem , Flavonoides/farmacocinética , Animais , Antineoplásicos/sangue , Disponibilidade Biológica , Carcinoma Pulmonar de Lewis/metabolismo , Carcinoma Pulmonar de Lewis/patologia , Linhagem Celular Tumoral , Ciclofosfamida/administração & dosagem , Feminino , Flavonoides/sangue , Flavonóis , Lipossomos , Camundongos , Camundongos Endogâmicos C57BL , Carga Tumoral/efeitos dos fármacosRESUMO
Pancreatic tumors are the gastrointestinal cancer with the worst prognosis in humans and with a survival rate of 5% at 5 years. Nowadays, no chemotherapy has demonstrated efficacy in terms of survival for this cancer. Previous study focused on the development of a new therapy by non thermal plasma showed significant effects on tumor growth for colorectal carcinoma and glioblastoma. To allow targeted treatment, a fibered plasma (Plasma Gun) was developed and its evaluation was performed on an orthotopic mouse model of human pancreatic carcinoma using a MIA PaCa2-luc bioluminescent cell line. The aim of this study was to characterize this pancreatic carcinoma model and to determine the effects of Plasma Gun alone or in combination with gemcitabine. During a 36 days period, quantitative BLI could be used to follow the tumor progression and we demonstrated that plasma gun induced an inhibition of MIA PaCa2-luc cells proliferation in vitro and in vivo and that this effect could be improved by association with gemcitabine possibly thanks to its radiosensitizing properties.
Assuntos
Antimetabólitos Antineoplásicos/farmacologia , Carcinoma Ductal Pancreático/terapia , Desoxicitidina/análogos & derivados , Neoplasias Pancreáticas/terapia , Gases em Plasma/uso terapêutico , Animais , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Rastreamento de Células , Terapia Combinada , Desoxicitidina/farmacologia , Modelos Animais de Doenças , Feminino , Humanos , Luciferases de Vaga-Lume/biossíntese , Camundongos , Neoplasias Pancreáticas/patologia , Carga Tumoral , Imagem Corporal Total , Ensaios Antitumorais Modelo de Xenoenxerto , Gencitabina , Neoplasias PancreáticasRESUMO
The natural flavonoid fisetin (3,3',4',7-tetrahydroxyflavone) has been shown to possess antiangiogenic and anticancer properties. Because of the limited water solubility of fisetin, our aim was to design and optimize a liposomal formulation that could facilitate its in vivo administration, taking into account the availability and cost of the various components. Several methods were evaluated such as probe sonication, homogeneization, film hydration and lipid cake formation. A selection of lipid and lipid-PEG was also performed via their incorporation in different formulations based on the size of the liposomes, their polydispersity index (PDI) and the fisetin encapsulation yield. An optimal liposomal formulation was developed with P90G and DODA-GLY-PEG2000, possessing a diameter in the nanometer scale (175nm), a high homogeneity (PDI 0.12) and a high fisetin encapsulation (73%). Fisetin liposomes were stable over 59 days for their particle diameter and still retained 80% of their original fisetin content on day 32. Moreover, liposomal fisetin retained the cytotoxicity and typical morphological effect of free fisetin in different tumour and endothelial cell lines. In conclusion, based on its physico-chemical properties and retention of fisetin biological effects, the developed liposomal fisetin preparation is therefore suitable for in vivo administration.