Celastrol enhances TRAIL-induced apoptosis in human glioblastoma via the death receptor pathway.

PURPOSE: Glioblastoma is the most common, malignant and devastating type of primary brain tumor. Tumor necrosis factor-related apoptosis-induced ligand (TRAIL) is characterized by its lethality to precancerous and cancerous cells. However, many kinds of tumor cells, including most glioma cells, tend to evade TRAIL-induced apoptosis. Celastrol is a pleiotropic compound from a traditional Chinese medicine that has proven to be useful as a sensitizer for TRAIL treatment. However, the underlying mechanism and role of celastrol in the sensitization of glioma cells remain to be elucidated. METHODS: The viability of glioma cell lines was examined by the CCK-8 assay. The expression of DR5 was detected by reverse transcriptase quantitative real-time PCR. The protein expression of DR5, cleaved caspase-8, cleaved caspase-3 and PARP were measured by western blot. The apoptosis rates and the sub-G1 population were detected by flow cytometry. The cellular morphological changes were assessed by TUNEL apoptosis and Hoechst 33258 staining assays. The knockdown of DR5 expression was conducted by siRNA. RESULTS: In this study, we observed that celastrol treatment inhibited cell viability in a dose-dependent manner, while glioma and normal human astroglial cell lines were resistant to TRAIL treatment. We also observed that the antiproliferative effects of TRAIL in combination with a noncytotoxic concentration of celastrol were significantly greater than those of celastrol or TRAIL alone. In addition, cell death induced by the combination treatment was apoptotic and occurred through the death receptor pathway via activation of caspase-8, caspase-3, and PARP. Furthermore, celastrol upregulated death receptor 5 (DR5) at the mRNA and protein levels, and siRNA-mediated DR5 knockdown reduced the killing effect of the combination drug treatment on glioma cells and reduced the activation of caspase-3, caspase-8 and PARP. CONCLUSIONS: Taken together, the results of our study demonstrate that celastrol sensitizes glioma cells to TRAIL via the death receptor pathway and that DR5 plays an important role in the effects of this cotreatment. The results indicate that this cotreatment is a promising tumor-killing therapeutic strategy with high efficacy and low toxicity.

Olfactory Ensheathing Cell-Conditioned Medium Reverts Aß25-35-Induced Oxidative Damage in SH-SY5Y Cells by Modulating the Mitochondria-Mediated Apoptotic Pathway.

Olfactory ensheathing cells (OECs) are a type of glia from the mammalian olfactory system, with neuroprotective and regenerative properties. ß-Amyloid peptides are a major component of the senile plaques characteristic of the Alzheimer brain. The amyloid beta (Aß) precursor protein is cleaved to amyloid peptides, and Aß25-35 is regarded to be the functional domain of Aß, responsible for its neurotoxic properties. It has been reported that Aß25-35 triggers reactive oxygen species (ROS)-mediated oxidative damage, altering the structure and function of mitochondria, leading to the activation of the mitochondrial intrinsic apoptotic pathway. Our goal is to investigate the effects of OECs on the toxicity of aggregated Aß25-35, in human neuroblastoma SH-SY5Y cells. For such purpose, SH-SY5Y cells were incubated with Aß25-35 and OEC-conditioned medium (OECCM). OECCM promoted the cell viability and reduced the apoptosis, and decreased the intracellular ROS and the lipid peroxidation. In the presence of OECCM, mRNA and protein levels of antioxidant enzymes (SOD1 and SOD2) were upregulated. Concomitantly, OECCM decreased mRNA and the protein expression levels of cytochrome c, caspase-9, caspase-3, and Bax in SH-SY5Y cells, and increased mRNA and the protein expression level of Bcl-2. However, OECCM did not alter intracellular Ca2+ concentration in SH-SY5Y cells. Taken together, our data suggest that OECCM ameliorates Aß25-35-induced oxidative damage in neuroblastoma SH-SY5Y cells by inhibiting the mitochondrial intrinsic pathway. These data provide new insights into the functional actions of OECCM on oxidative stress-induced cell damage.

Electrochemistry and electrogenerated chemiluminescence of a spirobifluorene-based donor (triphenylamine)-acceptor (2,1,3-benzothiadiazole) molecule and its organic nanoparticles.

A new D-A-π-A-D molecule (Spiro-BTA) containing two 2,1,3-benzothiadiazole (BTA) as the acceptor (A) and triphenylamine as the donor (D) bridged by a spirobifluorene moiety has been synthesized. The novel D-A molecule shows intense red emission (612 nm) with a high PL quantum yield (Φ(PL) = 0.51) in a solid film. A cyclic voltammogram of Spiro-BTA in 1:2 MeCN:benzene/0.1 M Bu(4)NPF(6) shows two reversible oxidation waves and one reversible reduction wave. The first oxidation wave and reduction wave were assigned as two successive electron transfer peaks separated by ∼50 mV related to the oxidation of the two noninteracting donors and the reduction of the two noninteracting acceptors, respectively. Electrogenerated chemiluminescence (ECL) of Spiro-BTA upon cyclic oxidation and reduction in MeCN:benzene 1:2 shows a very bright and stable red emission that could be seen in a well-lit room. Using a reprecipitation method, well-dispersed organic nanoparticles (NPs) of the Spiro-BTA were prepared in aqueous solution. The nanoparticles were analyzed by dynamic light scattering (DLS) and scanning electron microscopy (SEM), yielding a NP size (without surfactant) of 130 ± 20 nm, while with surfactant, 100 ± 20 nm. Bathochromic shifts of absorption spectra (∼16 ± 2 nm), as compared to that of the dissolved Spiro-BTA in THF, were observed for both NPs in water and as a thin film. While blue shifts (14 ± 2 nm) were observed for the photoluminescence (PL). The PL intensity of the Spiro-BTA nanoparticles was slightly enhanced (Φ(PL) of nanoparticles in water = 48%) over that of the dissolved Spiro-BTA in THF. The ECL of the organic Spiro-BTA nanoparticles in aqueous solution could be observed upon oxidation with tri-n-propylamine as a coreactant.