RESUMEN
Paeoniflorin-6'-O-benzene sulfonate (CP-25) has therapeutic potential for the treatment of hepatocellular carcinoma (HCC). 5-Fluorouracil (5-Fu) has been a conventional chemotherapeutic agent for HCC. Unfortunately, the nonspecific cytotoxicity and multidrug resistance caused by long-term use limited the clinical efficacy of 5-Fu. This study was aimed to investigate whether the combination of CP-25 and 5-Fu could generate synergistic effect in inhibiting HCC. The experiments on the diethylnitrosamine (DEN) -induced mice showed that compared with applying single drugs, the combination of CP-25 and 5-Fu presented stronger inhibition in tumor nodule and volume. Meanwhile, CP-25 and 5-Fu activated the intrinsic mitochondrial apoptosis pathway induced by P53, inhibited anti-apoptotic B-cell lymphoma (Bcl-2), induced the pro-apoptotic Bcl-2-associated X protein (Bax), Cytochrome-C and caspases. In addition, the synergistic effect was also validated in Bel-7402 and HepG-2 cells in vitro. This research not only provides a novel and effective combination strategy for the therapy of HCC but also provides an experimental basis for the development of CP-25 and 5-Fu compound preparation.
RESUMEN
Celastrol, an active ingredient of Tripterygium Wilfordii, is a traditional Chinese medicinal herb, which has attracted interests for its potential anti-inflammatory and anti-cancer activities. The aim of this study was to evaluate the anti-tumor effect of Celastrol against diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in rats and furthermore, to explore the underlying mechanism. Sprague-Dawley rats were intragastrically administered with DEN (10mg/kg) for 6 days every week and persisting 16 weeks. The number of nodules was calculated. Hematoxylin-Eosin (HE) staining was used to evaluate the hepatic pathological lesions. The levels of serum alanine aminotransferase (ALT), glutamic oxalacetic transaminase (AST), alkaline phosphatase (ALP) and alpha fetoprotein (AFP) were analyzed by Elisa kits, and the protein levels of p53, Murine double minute (MDM) 2, Bax, Bcl-2, Bcl-xl, cytochrome C, Caspase-3, Caspase-9 and Poly (ADP-ribose) polymerase (PARP) were analyzed by western blot. The results showed that Celastrol could significantly decrease the mortality, the number of tumor nodules and the index of liver in the Celastrol groups compared with DEN-treated group. Moreover, Celastrol obviously improved the hepatic pathological lesions and decreased the elevated levels of ALT, AST, ALP and AFP. Meanwhile, Celastrol suppressed the expression of the protein MDM2, activated the intrinsic mitochondrial apoptosis pathway induced by p53, inhibited anti-apoptotic Bcl-2 and Bcl-xl, induced the pro-apoptotic Bax, cytochrome C, PARP and caspases. These results suggested that Celastrol had a good therapeutic action in reversing DEN-induced HCC rats, which may be associated with the apoptosis of hepatoma cells induced by Celastrol.
Asunto(s)
Antineoplásicos/farmacología , Carcinoma Hepatocelular/inducido químicamente , Carcinoma Hepatocelular/prevención & control , Dietilnitrosamina/toxicidad , Neoplasias Hepáticas/inducido químicamente , Neoplasias Hepáticas/prevención & control , Triterpenos/farmacología , Animales , Carcinogénesis/efectos de los fármacos , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Citocromos c/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Mitocondrias/efectos de los fármacos , Triterpenos Pentacíclicos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Proteínas Proto-Oncogénicas c-mdm2/metabolismo , Ratas , Ratas Sprague-Dawley , Proteína p53 Supresora de Tumor/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Proteína bcl-X/metabolismoRESUMEN
Celastrol is a natural terpenoid isolated from Tripterygium wilfordii, a well-known Chinese medicinal herb that presents anti-proliferative activities in several cancer cell lines. Here, we investigated whether celastrol induces apoptosis on hepatocellular carcinoma Bel-7402 cells and further explored the underlying molecular mechanisms. Celastrol caused a dose- and time-dependent growth inhibition and apoptosis of Bel-7402 cells. It increased apoptosis through the up-regulation of Bax and the down-regulation of Bcl-2 in Bel-7402 cells. Moreover, celastrol induced the release of cytochrome c and increased the activation of caspase-3 and caspase-9, suggesting that celastrol-induced apoptosis was related to the mitochondrial pathway. These results indicated that celastrol could induce apoptosis in Bel-7402 cells, which may be associated with the activation of the mitochondria-mediated pathway.
Asunto(s)
Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Mitocondrias Hepáticas/genética , Mitocondrias Hepáticas/fisiología , Transducción de Señal/genética , Transducción de Señal/fisiología , Triterpenos/farmacología , Carcinoma Hepatocelular/genética , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Citocromos c/metabolismo , Relación Dosis-Respuesta a Droga , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Neoplasias Hepáticas/genética , Triterpenos Pentacíclicos , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Tripterygium/química , Triterpenos/aislamiento & purificación , Células Tumorales Cultivadas , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Dendritic cells (DCs) are the most powerful antigen-presenting cells that have an important role in the immunity and immune tolerance. Transforming growth factor ß (TGF-ß) is a pleiotropic cytokine widely expressing in various tissues and cells, which regulates cellular proliferation, differentiation and apoptosis of several immune cells and is considered to be a key factor in inducing immune tolerance. The effect of TGF-ß on DCs is very complex. In this study, we further investigated the effect of TGF-ß on inducing immune tolerance of DCs. DCs were differentiated from mice bone marrow cells in the absence or presence of TGF-ß. The phenotype as well as function was studied in detail. We found that TGF-ß limited the expression of CD40, CD83, CD86 and MHCII in DCs, increased CD45RB and indoleamine 2, 3-dioxygenase (IDO) expression in DCs, promoted IL-10 and limited IL-12 secretion by DCs. Moreover, TGF-ß increased the endocytosis ability of DCs and limited the ability of DCs in activating T cells. These results suggest that TGF-ß affects the immunity of DCs and enhances their tolerogenicity.