OBJECTIVETo study the effect of nano-liposome sustained elemene in inducing cell apoptosis of C6 glioma and to explore its influence on the expression of caspase-3 gene.

METHODSC6 glioma cells were cultured in medium with the same amount of nano-liposome sustained elemene and common elemene respectively, also in blank medium for control. The status of cell apoptosis was determined by flow cytometry at 0, 48 h and 72 h, and the expression of Caspase-3 protein was measured simultaneously by immunohistochemistry assay.

RESULTSMarked apoptosis presented in cells cultured in the medium with nano-liposome sustained elemene or common elemene at 48 h and 72 h, with the apoptotic rate significantly higher than that in the control. At the same time, Caspase-3 protein expression raised significantly in cells cultured in medium with either kinds of elemene, showing significant difference when compared with that in the control.

CONCLUSIONElemene has significant apoptosis promoting and Caspase-3 protein expression inducing effect on C6 glioma cells, which could be facilitated by nano-liposome bearing.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Gene Expression ; drug effects ; Glioma ; drug therapy ; enzymology ; genetics ; physiopathology ; Humans ; Liposomes ; Nanoparticles ; Plant Extracts ; pharmacology ; Sesquiterpenes ; pharmacology

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Animal ; Blotting, Northern/methods ; Brain/pathology ; Brain Neoplasms/pathology ; Brain Neoplasms/enzymology* ; Enzyme Activation ; Gelatinase A/metabolism ; Gelatinase A/genetics* ; Gelatinase B/metabolism ; Gene Expression Regulation, Enzymologic* ; Glioma/pathology ; Glioma/enzymology* ; Human ; Metalloendopeptidases/genetics* ; Papio ; Tissue Inhibitor-of Metalloproteinase-2/genetics ; Tissue-Inhibitor of Metalloproteinase-1/genetics ; Tumor Cells, Cultured

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Animal ; Blotting, Northern/methods ; Brain/pathology ; Brain Neoplasms/pathology ; Brain Neoplasms/enzymology* ; Enzyme Activation ; Gelatinase A/metabolism ; Gelatinase A/genetics* ; Gelatinase B/metabolism ; Gene Expression Regulation, Enzymologic* ; Glioma/pathology ; Glioma/enzymology* ; Human ; Metalloendopeptidases/genetics* ; Papio ; Tissue Inhibitor-of Metalloproteinase-2/genetics ; Tissue-Inhibitor of Metalloproteinase-1/genetics ; Tumor Cells, Cultured

The relevance of transglutaminases with neural function and several disorders has been emphasized recently. Especially, many polypeptides associated with neurodegenerative diseases are suggested to be putative transglutaminase substrates such as beta amyloid protein of Alzheimer's disease, microtubule-associated proteins and neurofilaments, etc. In addition, the CAG repeated gene products with probable polyglutamine tract, putative transglutaminase substrates, were identified in several neurodegenerative disorders. However, the identity of the brain transglutaminase has not been confirmed, because of enzymic stability and low activity. In the present experiment, we have isolated brain-specific transglutaminases, designated as TGase NI and TGase NII, which are different from other types of transglutaminases in respects of molecular weights (mw. 45 kDa, 29 kDa respectively), substrate affinity, elution profile on ion-exchange chromatography, sensitivity to proteases and ethanol, and immunological properties. The enzymes were localized specifically in the brain tissues but not in the liver tissue. And neural cells such as pheochromocytoma cell, glioma cell, primary neuronal and glial cells were shown to be enriched with TGase NI and TGase NII. The possible biological roles of the enzymes were discussed not only on the aspect of crosslinking activity but also of signal transducing capacity of the enzyme in the brain.
Animal ; Astrocytes/enzymology ; Blotting, Western ; Brain/enzymology* ; Calcium/metabolism ; Chromatography, Ion Exchange ; Endopeptidases/pharmacology ; Enzyme Stability ; Ethanol/pharmacology ; Glioma ; Immunoblotting ; Immunohistochemistry ; Male ; Molecular Weight ; Neurons/enzymology ; PC12 Cells ; Protein-Glutamine gamma-Glutamyltransferase/isolation & purification* ; Protein-Glutamine gamma-Glutamyltransferase/immunology ; Protein-Glutamine gamma-Glutamyltransferase/chemistry* ; Rats ; Rats, Sprague-Dawley ; Trypsin/pharmacology ; Tumor Cells, Cultured

OBJECTIVETo study the expression of Aurora-B in human glioma tissue and its significance.

METHODSThe total RNA was extracted from 41 human glioma tissues and 11 normal brain tissues by Trizol reagent. After reverse transcription of the total RNA into cDNAs, Aurora-B mRNA expressions in these samples were detected by quantitative real-time PCR. The protein expression in these samples was detected using immunohistochemical staining.

RESULTSAurora-B mRNA and protein expressions were significantly increased in glioma tissues as compared with those in normal brain tissues.

CONCLUSIONAurora-B mRNA and protein show markedly higher expressions in glioma tissue, suggesting that Aurora-B may be one of the malignant biomarkers in the pathogenesis and progression of human glioma.

Aurora Kinase B ; Aurora Kinases ; Biomarkers, Tumor ; metabolism ; Brain Neoplasms ; enzymology ; pathology ; Female ; Glioma ; metabolism ; pathology ; Humans ; Male ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo investigate the relationship between DNA-dependent protein kinase (DNA-PK) activity and anti-cancer drug sensitivity in human glioma tissues.

METHODSHuman glioma specimens were primarily cultured and its sensitivity to several anti-cancer drugs were evaluated by MTT assay. Nuclear protein was extracted from the glioma sample of the same patient and its DNA-PK activity was determined by a biotinylated DNA-PK assay with p53-derived peptide as a specific substrate.

RESULTSDNA-PK activity varied widely among these glioma samples. Of all 36 samples, 16 showed higher DNA-PK activity (relative activity > or = 0.40) and 20 samples with lower DNA-PK activity (relative activity < 0.40). The gliomas sensitive to DDP and VCR as evaluated by inhibition rate (IR > or = 50%) under plasma peak concentration (PPC) showed lower DNA-PK activity than the resistant ones (IR < 50%) (t = -3.445, P < 0.01). Furthermore, the gliomas with higher DNA-PK activity showed lower inhibition rate (IR < 50%) than those with lower DNA-PK activity ones (t = -2.145, P < 0.05).

CONCLUSIONDNA-PK activity is significantly associated with anti-cancer drug sensitivity to DDP and VCR in human gliomas. DNA-PK activity could be used as a new biomarker for the chemotherapy sensitivity of human gliomas.

Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cisplatin ; pharmacology ; DNA-Activated Protein Kinase ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Glioma ; enzymology ; pathology ; Humans ; Nuclear Proteins ; metabolism ; Vincristine ; pharmacology

OBJECTIVETo investigate the potentiality of herpes simplex virus thymidine kinase transduced endothelial progenitor cells (EPC-TK) as angiogenesis-targeting vector in the glioma treatment in vitro and in vivo.

METHODSEPC-TK were mixed with human umbilical vein endothelial cells (HUVECs), U87 or U251 cells at various ratios for ganciclovir (GCV) treatment. The bystander effect was observed by counting the survival cells using MTT assay, and the apoptotic cells were determined by annexin-V and propidium iodide (PI) staining. EPC-TK, EPCs, or phosphate buffered saline (PBS) were injected into the nude mice model of glioma xenograft by tail vein, for the EPC-TK group, EPC group, and PBS group, respectively. And then the changes of tumor volume and tumor vasculature were observed.

RESULTSGCV killed most EPC-TK and reduced the number of other viable cells in a cell:cell ratio-dependent and time-dependent manner. EPC-TK obviously inhibited tumor growth. The tumor volumes on day 21 were 1741.20+/- 576.10 mm(3), 3275.52 +/- 710.86 mm(3) and 3033.09+/-1134.86 mm(3) in the EPC-TK, EPC and PBS group, respectively. EPC-TK also displayed a significant effect on the inhibition of tumor angiogenesis.

CONCLUSIONEPC-TK can exert a potent antiglioma effect via inhibiting angiogenesis.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; Bystander Effect ; Cell Transformation, Viral ; physiology ; Endothelial Cells ; virology ; Endothelium ; Genetic Vectors ; Glioma ; therapy ; Humans ; Mice ; Mice, Nude ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; genetics ; Transduction, Genetic ; Transfection ; Xenograft Model Antitumor Assays

OBJECTIVETo study the anti-glioma activity of treatment by bone marrow stromal cells (BMSCs) transfected with AdCMV-tk containing HSV-tk gene in rats.

METHODSPrimary cultured BMSCs were obtained and transfected with HSV-tk (BMSCs/tk) and were injected into contralateral brain of glioma-bearing rats to observe their tropism for glioma cells. RT-PCR was performed to examine the transduct of tk gene after it was transduced into BMSCs. C6 glioma cells were co-cultured with BMSCs transfected with HSV-tk. MTT test was performed to examine its antitumor activity. BMSCs, after being transfected with HSV-tk, were injected into contralateral brain tissue of glioma-bearing rats to show their in vivo antitumor activity. Dynamic MRI was performed to monitor the development of intracranial glioma.

RESULTSPurified BMSCs were obtained by primary cultured bone marrow cells. After being transfected with HSV-TK, the cells still stably displayed extensive tropism for intracranial glioma and transcripted tk gene. RT-PCR showed that BMSCs/tk were transduced tk gene obviously at 21 days after AdCMV-tk transfection. BMSCs/tk showed a clear bystander effect after being co-cultured with C6 glioma cells in vitro. TUNEL assay showed that BMSCs/tk could obviously show bystander effect and induce apoptosis of glioma cells in vivo with an apoptosis positive ratio of 20.38% +/- 2.57%, showing a statistically significant difference in comparison with BMSCs group (2.56% +/- 0.52%, P = 0.023) and control group (2.74% +/- 0.38%, P = 0.025). Compared with the control group (21.40 +/- 1.63 days), BMSCs/tk transplantation significantly prolonged the survival time of glioma-bearing rats (52.60 +/- 13.11 days, P = 0.000). MRI detection showed that the least volume of intracranial glioma in BMSCs/tk group (8.28 +/- 2.64 mm3), significantly smaller than that in BMSCs group (134.51 +/- 16.37 mm3, P = 0.001) and control group (147.22 +/- 31.05 mm3, P = 0.001). Some of the intracranial gliomaa disappeared after transplantation of BMSCs/tk.

CONCLUSIONBMSCs transfected with AdCMV-tk may become an effective therapy method in the treatment for glioma.

Animals ; Apoptosis ; Bone Marrow Cells ; cytology ; Brain ; pathology ; Bystander Effect ; Cell Line, Tumor ; Coculture Techniques ; Genetic Therapy ; methods ; Glioma ; pathology ; therapy ; Magnetic Resonance Imaging ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Simplexvirus ; enzymology ; Stromal Cells ; cytology ; enzymology ; transplantation ; Thymidine Kinase ; genetics ; metabolism ; Transfection

OBJECTIVETo explore the role of phospholipase C-gamma1 (PLC-gamma1) in tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis of human glioma SWO cells.

METHODSThe PLC-gamma1 pathway was blocked by U73122 in SWO cells, and the inhibitory effect of TNF-alpha on SWO glioma cell proliferation with or without U73122 treatment was investigated by MTT assay. The cell apoptosis induced by TNF-alpha along or in combination with U73122 was detected by flow cytometry with PI staining. The expression of caspase-3 and Bcl-2 was detected by Western blotting.

RESULTS AND CONCLUSIONU73122 can sensitize SWO glioma cells to TNF-alpha-induced apoptosis. Blocking the PLC-gamma1 pathway may not induce apoptosis of SWO glioma cells, but can sensitize SWO glioma cells to small-dose TNF-alpha-induced apoptosis, the mechanism of which may involve down-regulation of bcl-2.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Estrenes ; pharmacology ; Flow Cytometry ; Glioma ; enzymology ; pathology ; Humans ; Phosphodiesterase Inhibitors ; pharmacology ; Phospholipase C gamma ; antagonists & inhibitors ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrrolidinones ; pharmacology ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology