RESUMEN
The purpose of these experiments was to identify growth factors produced during the formation of a peritoneal wound in relation to tumour cell seeding and stimulated growth in granulation tissue. Gelfoam(c) gelatin sponge was implanted in the mesenteric fan of nude mice to initiate the granulation process. Human HT29 colon carcinoma cells were inoculated intraperitoneally at various times after sponge implantation and tumour growth in granulation tissue was determined. RNA isolated from granulation tissue was used for polymerase chain reaction analysis of the expression of specific growth factors and receptors [vascular endothelial growth factor (VEGF), transforming growth factor-beta (TGF-beta) and lysophosphatic acid (LPA)], and for microarray analysis of differentially expressed genes in early vs. late granulation tissue. Inflammatory cells infiltrated the sponge within 1 day, followed by fibroblasts and the formation of an extracellular matrix. Tumour cell inoculation at 8 h to 3 days after sponge implantation resulted in extensive tumour formation in all cases. Inoculation at 10-28 days resulted in focal tumour growth in only 16-33% of the sponges. Low amounts of VEGF, TGF-beta(1-3), TGF-beta RIII and LPA receptors 1,2 were detected in early granulation tissue, with increased expression from day 10. Microarray analysis identified additional differentially expressed genes that may stimulate tumour take and growth in early granulation tissue.
Asunto(s)
Tejido de Granulación/metabolismo , Sustancias de Crecimiento/análisis , Siembra Neoplásica , Cicatrización de Heridas , Animales , Femenino , Gelatina , Expresión Génica , Células HT29 , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Análisis de Secuencia por Matrices de Oligonucleótidos , Peritoneo/lesiones , Receptores Acoplados a Proteínas G/análisis , Receptores del Ácido Lisofosfatídico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Crecimiento Transformador beta/análisis , Factor A de Crecimiento Endotelial Vascular/análisis , Cicatrización de Heridas/genéticaRESUMEN
Animal tumour models using orthotopic tumours for the evaluation of cancer therapies are of greater clinical relevance than subcutaneous models, but they also pose greater difficulties for measuring tumour size and quantifying response to treatment. In this study, we used noninvasive bioluminescence imaging to monitor the intraperitoneal growth of luciferase-transfected CC531 colorectal cells in adult WAG/RIJ rats. The bioluminescence signal correlated well with post-mortem assessment of tumour load by visual inspection of the peritoneal cavity at specific follow-up times. Using bioluminescence imaging, we were able to monitor peritoneal tumour growth sequentially in time and to calculate a tumour growth rate for each animal; this is not possible with invasive methods of evaluating tumour load. Bioluminescence imaging of rats treated with a single dose of cisplatin (4 mg x kg(-1), i.p.) demonstrated a significant delay in peritoneal tumour growth relative to saline controls (mean 45.0+/-s.d. 13.0 vs 28.2+/-10.3 days; P=0.04). Similar protocols evaluated by visual scoring of tumour load at 40 days after inoculation supported these findings, although no quantitative assessment of treatment-induced growth delay could be made by this method. This study shows that in vivo imaging of luciferase-transfected tumour cells is a useful tool to investigate the dynamics of disseminated tumour growth and efficacy of anticancer treatment in orthotopic models of peritoneal cancer in rats. It offers an attractive alternative to invasive methods, and requires fewer animals for measuring tumour response to therapy.
Asunto(s)
Carcinoma/patología , Neoplasias del Colon/patología , Mediciones Luminiscentes , Neoplasias Experimentales , Animales , Carcinoma/tratamiento farmacológico , Carcinoma/veterinaria , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/veterinaria , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Luciferasas/análisis , Luciferasas/genética , Neoplasias Peritoneales , Ratas , Transfección , Resultado del TratamientoRESUMEN
BACKGROUND: HIPEC is a new treatment modality for abdominal cancers that combines cytoreductive surgery with Hyperthermic, Intraoperative Peritoneal Chemotherapy, followed by systemic chemotherapy. A significant survival benefit has been shown for HIPEC compared with systemic therapy alone. However, it is not clear what is the contribution of i.p. drug delivery and what influence the mild hyperthermia has on the uptake of cisplatin in abdominal tumors. MATERIALS AND METHODS: We used a peritoneal perfusion system in rats to compare the pharmacokinetics and pharmacodynamics of cisplatin, after normothermic (37 degrees C/90 minutes) and hyperthermic (40 degrees C/90 minutes) intra-peritoneal perfusion, with an i.p. bolus injection. RESULTS: Hyperthermic perfusion with 15 micrograms/ml (in 200 ml) cisplatin gave equivalent plasma drug levels to a maximum tolerated dose (MTD) i.p. bolus injection of 4 mg/kg (36 micrograms/ml in 20 ml). The drug concentration in small (1-5 mm) intra-peritoneal tumors was also comparable for both these treatments, and for normothermic perfusion. CONCLUSION: Mild hyperthermic perfusion with cisplatin (40 degrees C/90 minutes) did not improve drug uptake in small intra-peritoneal tumors, relative to normothermic perfusion or i.p. bolus injection.
Asunto(s)
Antineoplásicos/farmacología , Cisplatino/farmacología , Hipertermia Inducida , Neoplasias Peritoneales/terapia , Absorción , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/sangre , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Cisplatino/administración & dosificación , Cisplatino/sangre , Cisplatino/farmacocinética , Cisplatino/uso terapéutico , Neoplasias del Colon/patología , Terapia Combinada , Femenino , Infusiones Parenterales , Inyecciones Intraperitoneales , Periodo Intraoperatorio , Riñón/metabolismo , Hígado/metabolismo , Dosis Máxima Tolerada , Trasplante de Neoplasias , Perfusión , Neoplasias Peritoneales/tratamiento farmacológico , Neoplasias Peritoneales/secundario , Peritoneo/metabolismo , Ratas , Distribución TisularRESUMEN
Based on observations that the persistent environmental pollutant hexachlorobenzene (HCB) induces inflammatory skin lesions and eosinophilic and granulomatous lung pathology as well as in vivo airways hyperresponsiveness to methacholine in the BN/SsNOlaHsd rat (Michielsen et al., Toxicol Appl Pharmacol 172:11-20, 2001), which are features of human Churg-Strauss syndrome (CSS), we have investigated whether HCB induced other features of CSS such as asthma and systemic vasculitis involving the heart and kidneys in this strain of rat. To this end, BN/SsNOlaHsd rats received control feed or feed supplemented with 450 mg/kg HCB. On days 6, 14 or 21, tracheas were isolated to assess non-specific in vitro airways hyperresponsiveness (AHR) to cumulative concentrations of arecoline and serotonin. In addition, lungs were lavaged to count and differentiate lavage cells, and skin, lungs, heart, kidneys, and lymph nodes were processed for histopathological investigation. HCB induced eosinophilic and granulomatous lung pathology in the BN/SsNOlaHsd rat, which became more severe with time and was associated with significant in vitro AHR to arecoline. Moreover, as in CSS-patients, systemic effects on spleen and lymph nodes were observed in HCB-fed BN/SsNOlaHsd rats, as well as development of skin lesions with vascular changes and eosinophilic infiltrates. In contrast, cardiac or renal involvement, frequently seen in CSS-patients, was not seen in HCB-fed rats. More importantly, there were no indications of necrotizing vasculitis, a hallmark feature of CSS, in the lungs and skin of BN/SsNOlaHsd rats. Thus, it is concluded that the persistent environmental pollutant HCB possibly induces a mild or early stage of CSS in the BN/SsNOlaHsd rat that may evolve into fully developed CSS after prolonged exposure to HCB.
Asunto(s)
Hiperreactividad Bronquial/inducido químicamente , Síndrome de Churg-Strauss/inducido químicamente , Contaminantes Ambientales/toxicidad , Fungicidas Industriales/toxicidad , Hexaclorobenceno/toxicidad , Eosinofilia Pulmonar/inducido químicamente , Animales , Arecolina/farmacología , Hiperreactividad Bronquial/patología , Líquido del Lavado Bronquioalveolar/citología , Síndrome de Churg-Strauss/patología , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunoglobulina E/sangre , Técnicas In Vitro , Riñón/patología , Pulmón/efectos de los fármacos , Pulmón/patología , Ganglios Linfáticos/patología , Modelos Animales , Miocardio/patología , Eosinofilia Pulmonar/patología , Ratas , Ratas Endogámicas BN , Serotonina/farmacología , Piel/efectos de los fármacos , Piel/patologíaRESUMEN
The urokinase-type plasminogen activator (uPA) system plays an important role in tumor cell invasion, metastases, and angiogenesis. uPA, uPA receptor, and plasminogen activator inhibitor 1 (PAI-1) are prognostic factors in different solid tumors, e.g., renal cell carcinomas (RCCs). von Hippel-Lindau (VHL) disease is an inherited cancer syndrome that is characterized by extensively vascularized tumors, including hemangioblastomas and RCCs. In 75% of sporadic RCCs, the VHL gene is also inactivated. It has been recognized in sporadic RCC that PAI-1 mRNA levels are up-regulated and uPA mRNA levels are down-regulated. We determined the role of the VHL tumor suppressor gene in the regulation of the uPA system in RCC. In 786-O RCC cells expressing the wild-type (wt) VHL gene, we measured a 3-fold higher overall urokinase activity than in 786-O cells expressing a mutant VHL gene or lacking VHL. uPA mRNA and protein levels were higher in cells with wt VHL compared with cells with mutant VHL or lacking VHL. In addition, PAI-1 mRNA and protein levels were dramatically increased in 786-O cells with mutant VHL or lacking VHL, compared with cells expressing wt VHL. Our results provide further evidence that the VHL gene plays an important role in the process of angiogenesis by regulation of plasmin-mediated proteolysis of the extracellular matrix and may explain why VHL-induced RCCs grow slowly and metastasize relatively late.