ABSTRACT
Tumor necrosis factor-related apoptosis-induced ligand (TRAIL) induces apoptosis selectively in cancer cells while sparing normal cells. However, many cancer cells are resistant to TRAIL-induced cell death. Here, we report that paxilline, an indole alkaloid from Penicillium paxilli, can sensitize various glioma cells to TRAIL-mediated apoptosis. While treatment with TRAIL alone caused partial processing of caspase-3 to its p20 intermediate in TRAIL-resistant glioma cell lines, co-treatment with TRAIL and subtoxic doses of paxilline caused complete processing of caspase-3 into its active subunits. Paxilline treatment markedly upregulated DR5, a receptor of TRAIL, through a CHOP/GADD153-mediated process. In addition, paxilline treatment markedly downregulated the protein levels of the short form of the cellular FLICE-inhibitory protein (c-FLIPS) and the caspase inhibitor, survivin, through proteasome-mediated degradation. Taken together, these results show that paxilline effectively sensitizes glioma cells to TRAIL-mediated apoptosis by modulating multiple components of the death receptor-mediated apoptotic pathway. Interestingly, paxilline/TRAIL co-treatment did not induce apoptosis in normal astrocytes, nor did it affect the protein levels of CHOP, DR5 or survivin in these cells. Thus, combined treatment regimens involving paxilline and TRAIL may offer an attractive strategy for safely treating resistant gliomas.
Subject(s)
Humans , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Astrocytes/metabolism , CASP8 and FADD-Like Apoptosis Regulating Protein/genetics , Caspase 3/metabolism , Cell Line, Tumor , Drug Discovery , Flow Cytometry , Glioma/metabolism , Indoles/pharmacology , Inhibitor of Apoptosis Proteins/metabolism , RNA, Small Interfering , Receptors, TNF-Related Apoptosis-Inducing Ligand/genetics , Reverse Transcriptase Polymerase Chain Reaction , TNF-Related Apoptosis-Inducing Ligand/metabolism , Transcription Factor CHOP/analysisABSTRACT
A recent study reported that a diet rich in isoflavones is beneficial for bone formation in growing rats. It therefore seemed desirable to find out whether the beneficial effect of isoflavones in ovariectomized rats could also be reproduced with same amount of isoflavones which used for growing rats. To study the effect of isoflavones, an equal amount of isoflavones which used for growing rats, on bone mineral density and bone mineral content in ovariectmized rats were performed. Forty female Sprague-Dawley rats (body weight 210+/-5 g) were divided into two groups, ovariectomy and sham groups, which were each randomly divided into two subgroups that were fed casein and casein supplemented with isoflavones diets for 9 weeks after operation. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin) in spine and femur. Serum alkaline phosphatase activity (ALP) and osteocalcin and urinary DPD crosslinks value were measured as markers of bone formation and resorption. The results of this study indicate that body weight gain and food intake were higher in ovariectomy groups than in sham groups regardless of diets. Serum Ca concentration was lower in ovariectomy groups than in SHAM groups. Serum ALP, osteocalcin, and crosslink value were increased in ovariectomy groups. Spine BMD/weight, femur BMD/weight, and femur BMC/weight of ovariectomy groups were significantly lower than SHAM groups after 9 weeks. However, isoflavones supplemented group in ovariectomy groups, serum ALP and osteocalcin concentrations, spine BMD/weight and spine BMC, femur BMD/weight and femur BMC/weight were significantly increased after 9 weeks. In conclusion, the beneficial effect of isoflavones on bone in ovareiectomized rats was shown on 9 weeks after feeding with an equal amount of isoflavones supplementation which used for growing rats.