RESUMEN
Balanites aegyptiaca is a widely distributed African plant of medicinal interest containing a number of cytotoxic and cytostatic compounds. The studies reported here have attempted to further characterize the anti-cancer activity of a mixture of steroidal saponins: balanitin-6 (28%) and balanitin-7 (72%) isolated from Balanites aegyptiaca kernels. The balanitin-6 and -7 mixture (henceforth referred to as bal6/7) has demonstrated appreciable anti-cancer effects in human cancer cell lines in vitro. Bal6/7 displayed higher anti-proliferative activity than etoposide and oxaliplatin, although the mixture was appreciably less active than SN38 and markedly less active than taxol. Bal6/7 demonstrated highest activity against A549 non-small cell lung cancer (NSCLC) (IC(50), 0.3 microM) and U373 glioblastoma (IC(50), 0.5 microM) cell lines. The current study has further indicated that bal6/7 is more a cytotoxic compound than a cytostatic one. However, Bal6/7 does not appear to mediate its anti-proliferative effects by inducing apoptotic cell death. Computer-assisted cellular imaging has revealed that bal6/7 does not induce detergent-like effects in A549 NSCLC and U373 glioblastoma unlike certain saponins. Furthermore there is indication that its in vitro anti-cancer activities result at least partly from depletion of [ATP]i, leading in turn to major disorganization of actin cytoskeleton, ultimately resulting in the impairment of cancer cell proliferation and migration. In contrast to a number of natural products acting as anti-cancer agents, bal6/7 does not induce an increase in intra-cellular reactive oxygen species. In vivo, bal6/7 increased the survival time of mice bearing murine L1210 leukemia grafts to the same extent reported for vincristine. These preliminary in vivo data suggest that it may be possible to generate novel hemi-synthetic derivatives of balanitin-6 and -7 with potentially improved in vitro and in vivo anti-cancer activity and reduced in vivo toxicity, thus markedly improving the therapeutic ratio.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Balanites/química , Diosgenina/análogos & derivados , Diosgenina/farmacología , Saponinas/farmacología , Adenosina Trifosfato/metabolismo , Animales , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Diosgenina/uso terapéutico , Femenino , Humanos , Leucemia L1210/tratamiento farmacológico , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Especies Reactivas de Oxígeno/metabolismo , Saponinas/toxicidadRESUMEN
Our study has shown that the Amaryllidaceae isocarbostyril narciclasine induces marked apoptosis-mediated cytotoxic effects in human cancer cells but not in normal fibroblasts by triggering the activation of the initiator caspases of the death receptor pathway (caspase-8 and caspase-10) at least in human MCF-7 breast and PC-3 prostate carcinoma cells. The formation of the Fas and death receptor 4 (DR4) death-inducing signaling complex was clearly evidenced in MCF-7 and PC-3 cancer cells. Caspase-8 was found to interact with Fas and DR4 receptors on narciclasine treatment. However, narciclasine-induced downstream apoptotic pathways in MCF-7 cells diverged from those in PC-3 cells, where caspase-8 directly activated effector caspases such as caspase-3 in the absence of any further release of mitochondrial proapoptotic effectors. In contrast, in MCF-7 cells, the apoptotic process was found to require an amplification step that is mitochondria-dependent, with Bid processing, release of cytochrome c, and caspase-9 activation. It is postulated that the high selectivity of narciclasine to cancer cells might be linked, at least in part, to this activation of the death receptor pathway. Normal human fibroblasts appear approximately 250-fold less sensitive to narciclasine, which does not induce apoptosis in these cells probably due to the absence of death receptor pathway activation.
Asunto(s)
Alcaloides de Amaryllidaceae/farmacología , Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/patología , Carcinoma/patología , Mitocondrias/fisiología , Fenantridinas/farmacología , Neoplasias de la Próstata/patología , Receptores del Factor de Necrosis Tumoral/fisiología , Alcaloides de Amaryllidaceae/aislamiento & purificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/fisiología , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/fisiología , Neoplasias de la Mama/metabolismo , Carcinoma/metabolismo , Caspasas/fisiología , Citocromos c/análisis , Fragmentación del ADN , Resistencia a Antineoplásicos , Activación Enzimática/efectos de los fármacos , Femenino , Fibroblastos/efectos de los fármacos , Humanos , Masculino , Mitocondrias/enzimología , Narcissus/química , Proteínas de Neoplasias/efectos de los fármacos , Proteínas de Neoplasias/fisiología , Fenantridinas/aislamiento & purificación , Neoplasias de la Próstata/metabolismo , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF , Receptor fas/efectos de los fármacos , Receptor fas/fisiologíaRESUMEN
Amonafide (1), a naphthalimide which binds to DNA by intercalation and poisons topoisomerase IIalpha, has demonstrated activity in phase II breast cancer trials, but has failed thus far to enter clinical phase III because of dose-limiting bone marrow toxicity. Compound 17 (one of 41 new compounds synthesized) is a novel anticancer naphthalimide with a distinct mechanism of action, notably inducing autophagy and senescence in cancer cells. Compound 17 (2,2,2-trichloro-N-({2-[2-(dimethylamino)ethyl]-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinolin-5-yl}carbamoyl)acetamide (UNBS3157)) was found to have a 3-4-fold higher maximum tolerated dose compared to amonafide and not to provoke hematotoxicity in mice at doses that display significant antitumor effects. Furthermore, 17 has shown itself to be superior to amonafide in vivo in models of (i) L1210 murine leukemia, (ii) MXT-HI murine mammary adenocarcinoma, and (iii) orthotopic models of human A549 NSCLC and BxPC3 pancreatic cancer. Compound 17, therefore, merits further investigation as a potential anticancer agent.
Asunto(s)
Acetamidas/síntesis química , Antineoplásicos/síntesis química , Isoquinolinas/síntesis química , Naftalimidas/síntesis química , Urea/análogos & derivados , Acetamidas/farmacología , Acetamidas/toxicidad , Adenina , Animales , Antineoplásicos/farmacología , Antineoplásicos/toxicidad , Apoptosis , Autofagia , Camptotecina/análogos & derivados , Camptotecina/farmacología , Línea Celular Tumoral , Senescencia Celular , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Recuento de Eritrocitos , Femenino , Humanos , Imidas/farmacología , Imidas/toxicidad , Irinotecán , Isoquinolinas/farmacología , Isoquinolinas/toxicidad , Recuento de Leucocitos , Dosis Máxima Tolerada , Ratones , Naftalimidas/farmacología , Naftalimidas/toxicidad , Trasplante de Neoplasias , Organofosfonatos , Recuento de Plaquetas , Relación Estructura-Actividad , Inhibidores de Topoisomerasa I , Urea/síntesis química , Urea/farmacología , Urea/toxicidad , GemcitabinaRESUMEN
BACKGROUND: The Helicase-Like Transcription Factor (HLTF/SMARCA3) belongs to the family of SWI/SNF proteins that use the energy of ATP hydrolysis to remodel chromatin in a variety of cellular processes. Several SWI/SNF genes are disrupted in cancer, suggesting a role of tumor suppressor. Similarly, the HLTF gene was recently found to be inactivated by hypermethylation in a number of advanced colon and gastric tumors. However, other evidences indicated a 20-fold HLTF overexpression in cell lines derived from various neoplasms (ovary, breast, cervix, kidney...). RESULTS: In the present study, we investigated HLTF expression by immunohistochemistry in a model of kidney tumors induced by continuous administration of diethylstilbestrol to male Syrian golden hamsters. A strong labeling was already detected in small tumor buds, making HLTF an early cancer marker in this model. Although every cell stained for HLTF at this early stage, the number of HLTF-positive cells decreased to 10% with cancer progression, and these positive cells were dispersed in the tumor mass. HLTF expression was conserved in the HKT-1097 cell line established from kidney tumors, but again only 10% of positive cells were found in xenografts produced by HKT-1097 cells in nude mice. CONCLUSION: In conclusion, our data suggest that HLTF gene activation is linked to initial steps of carcinogenesis in this model and should be investigated in early stages of other neoplasms.
Asunto(s)
ADN Helicasas/metabolismo , Proteínas de Unión al ADN/metabolismo , Estrógenos/farmacología , Neoplasias Renales/genética , Factores de Transcripción/metabolismo , Animales , Secuencia de Bases , Cricetinae , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/genética , Dietilestilbestrol/farmacología , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Inmunohistoquímica , Riñón/efectos de los fármacos , Riñón/metabolismo , Riñón/patología , Neoplasias Renales/inducido químicamente , Neoplasias Renales/metabolismo , Masculino , Mesocricetus , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Neoplasias Experimentales/genética , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Homología de Secuencia de Ácido Nucleico , Factores de Transcripción/análisis , Factores de Transcripción/genética , Activación Transcripcional , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
Non-small cell lung cancers (NSCLCs) are the leading cause of cancer deaths in most developed countries. Targeting heat shock protein 70 (Hsp70) expression and function, together with the induction of lysosomal membrane permeabilization (LMP), could overcome the multiple anti-cell death mechanisms evidenced in NSCLCs that are responsible for the failure of currently used chemotherapeutic drugs. Because cardenolides bind to the sodium pump, they affect multiple signaling pathways and thus have a number of marked effects on tumor cell behavior. The aim of the present study was to characterize in vitro and in vivo the antitumor effects of a new cardenolide (UNBS1450) on experimental human NSCLCs. UNBS1450 is a potent source of in vivo antitumor activity in the case of paclitaxel-and oxaliplatin-resistant subcutaneous human NCI-H727 and orthotopic A549 xenografts in nude mice. In vitro UNBS1450-mediated antitumor activity results from the induction of nonapoptotic cell death. UNBS1450 mediates the decrease of Hsp70 at both mRNA and protein levels, and this is at least partly due to UNBS1450-induced downregulation of NFAT5/TonEBP (a factor responsible for the transcriptional control of Hsp70). These effects were paralleled by the induction of LMP, as evidenced by acridine orange staining and immunofluorescence analysis for cathepsin B accumulation.
Asunto(s)
Antineoplásicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Cardenólidos/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Lisosomas/química , Naranja de Acridina/farmacología , Animales , Catepsina B/metabolismo , Línea Celular Tumoral , Modelos Animales de Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica , Proteínas HSP70 de Choque Térmico/metabolismo , Humanos , Ratones , Ratones DesnudosRESUMEN
The developmental competence of bovine oocytes isolated from antral follicles of different sizes was assessed in three European laboratories (Belgium, UCL; Denmark, DIAS; France, INRA). Using the same protocol for in vitro production of embryos, the oocytes isolated from follicles with a diameter > or = 6 mm always gave a higher blastocyst rate than oocytes from follicles < 4 mm (UCL: 42% versus 14%, DIAS: 50% versus 35%, INRA: 39% versus 22%; P < 0.05). Blastocyst cell number was not affected by follicle size. Several parameters were investigated for these oocytes. The energy metabolism of cumulus-oocyte-complexes and of denuded oocytes was assessed by the oxygen and pyruvate uptake and by lactate release both at the beginning and the end of the maturation. No effect of follicle size could be detected but lactate release increased after maturation. The global profile of transcripts, the pattern of protein neosynthesis and the kinetics of meiosis resumption were not affected by follicle size. The developmental kinetics of derived embryos was also analysed. Whatever the follicle size, viable embryos had a shorter first and third embryonic cell cycle. Among the viable embryos, the size of the follicle interfered with the fourth cell cycle duration. A higher percentage of blastocysts issued from large follicle presented a short fourth cell cycle (9h) (35% versus 6%; P < 0.05). Beside, blastocysts derived from small follicles had a delayed cavitation and expansion. Thereby, a higher developmental competence for oocytes from follicle > or = 6 mm versus < 4 mm was demonstrated in three laboratories although no differences could be displayed directly at the oocyte level.