OBJECTIVE The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS First,the model of C6 glioma in Wistar rats w...OBJECTIVE The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS First,the model of C6 glioma in Wistar rats was established and verified by hemotoxylin and eosin staining,immunohistochemical staining and magnetic resonance imaging(MRI).Then different doses of lapachol were gavaged and tumor volumes of the C6 glioma were detected by MRI.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phen-azinemethosulfate(PMS)assay,Hoechst33358 staining,AnnexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomeraseⅠ(TOPⅠ)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322 DNA relaxation assay.Molecular docking was used to predict the interaction of lapachol-TOPⅠand lapachol-TOPⅡ.TOP I and TOPⅡexpression levels in C6 cells were determined by Enzymelinked immunosorbent assay kits and real-time polymerase chain reaction(RT-PCR).RESULTS The rat C6 glioma model was successfully established.High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).MTS/PMS assay,Hoechst 33258 staining,AnnexinⅤ-FITC/PI staining,and comet assay showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cells in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠandⅡ.Molecular docking showed that lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡ.Enzyme-linked immunosorbent assay and RT-PCR showed that lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠand TOPⅡactivities,as wel as TOPⅡexpression.展开更多
Successful chemotherapy with paclitaxel(PTX)is impeded by multidrug resistance(MDR)in tumor cells.In this study,lipid-albumin nanoassemblies co-loaded with borneol and paclitaxel(BOR/PTX LANs)were prepared to circumve...Successful chemotherapy with paclitaxel(PTX)is impeded by multidrug resistance(MDR)in tumor cells.In this study,lipid-albumin nanoassemblies co-loaded with borneol and paclitaxel(BOR/PTX LANs)were prepared to circumvent MDR in C6 glioma cells.The physiochemical properties including particle size,encapsulation efficiency and morphology were evaluated in vitro.Quantitative and qualitative investigations of cellular uptake were carried out in C6 glioma cells.The cytotoxicity of the BOR/PTX LANs was determined by MTT assay.After that,the tumor targeting was also evaluated in C6 glioma bearing mice by in vivo imaging analysis.BOR/PTX LANs have a higher entrapment efficiency(90.4±1.2%),small particle size(107.5±3.2 nm),narrow distribution(P.I.=0.171±0.02).The cellular uptake of PTX was significantly increased by BOR/PTX LANs compared with paclitaxel loaded lipidalbumin nanoassemblies(PTX LANs)in quantitative research.The result was further confirmed by confocal laser scanning microscopy qualitatively.The cellular uptake was energy-,timeand concentration-dependent,and clathrin-and endosome/lysosome-associated pathways were involved.The BOR/PTX LANs displayed a higher cytotoxicity agaist C6 glioma cells in comparion with PTX LANs and Taxol.Moreover,the encapsulation of BOR in LANs obviously increased the accumulation of the drug in tumor tissues,demonstrating the tumor targeted ability of BOR/PTX LANs.These results indicated that BOR/PTX LANs could overcome MDR by combination of drug delivery systems and P-gp inhibition,and shown the potential for treatment of gliomas.展开更多
BACKGROUND: Embryonic neural stem cells (NSCs) have provided positive effects for the treatment of glioma. However, the source for embryonic NSCs remains limited and high amplification conditions are required. Bone ma...BACKGROUND: Embryonic neural stem cells (NSCs) have provided positive effects for the treatment of glioma. However, the source for embryonic NSCs remains limited and high amplification conditions are required. Bone marrow stromal cells (BMSCs) have been proposed for the treatment of glioma. OBJECTIVE: To investigate biological changes in NSCs and BMSCs following transplantation into rat models of glioma. DESIGN, TIME AND SETTING: A randomized, controlled, animal experiment was performed at the Embryonic Stem Cell Research Laboratory of Yunyang Medical College from February 2006 to August 2008. MATERIALS: The rat C6 glioma cell line was purchased from Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; mouse anti-bromodeoxyuridine (BrdU) monoclonal antibody and Cy3-labeled goat anti-mouse IgG antibody was purchased from Upstate, USA. METHODS: A total of 95 Sprague Dawley rats were randomly assigned to three groups: NSC (n = 35), transplanted with > 6 × 106 NSCs via left medial hind limb; BMSC (n = 35), transplanted with > 1 × 106 BMSCs via left medial hind limb; model group (n = 25), injected with the same volume of 0.1 mmol/L phosphate buffered saline. MAIN OUTCOME MEASURES: Gliomal growth and size were assessed by nuclear magnetic resonance, and glioma morphological features were observed following hematoxylin-eosin staining and BrdU immunohistochemistry 3 and 4 weeks following transplantation. RESULTS: The average survival of rats in the BMSC, NSC, and model groups was 4.03, 4.28, and 3.88 weeks. At 3 weeks, there was no significant difference in the average glioma diameter between the BMSC and model groups (P > 0.05). However, gliomal diameter was significantly decreased in the NSC group compared with the model group (P < 0.05). At 4 weeks, there was no statistical difference between the groups (P > 0.05). BrdU immunohistochemistry revealed that BMSCs and NSCs appeared to migrate to the gliomas. CONCLUSION: NSCs inhibited glioma cell growth and prolonged rat survival. BMSCs did not significantly suppress glioma cell growth.展开更多
OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in v...OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phenazinemethosulfate(PMS)assay,hoechst 33358 staining,annexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomerase I(TOP I)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322DNA relaxation assays and molecular docking.TOPⅠand TOPⅡexpression levels in C6 cells were also determined.RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).It was showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cel s in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠ and Ⅱ.Lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡin molecular docking study.Also,lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠ and TOPⅡ activities,as wel as TOPⅡ expression.展开更多
Objective To investigate the anti-tumor effect and mechanism of tamoxifen on rat C6 glioma cells. Methods C6 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 3% fetal calf serum (FCS), and...Objective To investigate the anti-tumor effect and mechanism of tamoxifen on rat C6 glioma cells. Methods C6 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 3% fetal calf serum (FCS), and treated with tamoxifen of different concentrations, i.e. group A (1.25μmol/L), group B (2.50 μmol/L), group C (5.00 μmol/L), group D (10.00 μmol/L), group E (20.00 μmol/L) and control group (0.00 μmol/L). Morphological changes, MTT assay and 5-bromo-2'-deoxyuriding labeling ratio were assessed. Apoptosis was observed by flow cytometry. Results C6 cells treated with different doses of tamoxifen for 24, 48, and 72 hours became irregular in shape, while cells treated with vehicle grew normally. MTT assay showed that tamoxifen did not suppress C6 cell growth until 72 hours after treatment. Seventy-two hours after treatment, there were significant differences in cell viable rate between group A versus groups C, D and E; so did group B versus group D as well as group E (P<0.05). BrdU incorporation assay indicated significant difference of BrdU labbled index (BrdU LI) among groups A, C, E and control group 48 hours after treatment (P<0.05). And the BrdU LI decreased with the increased concentration of tamoxifen. Flow cytometry (FCM) showed significant difference between treated group and control group at 24, 48, and 72 hours after treatment (P<0.05). Conclusion Tamoxifen significantly suppresses the growth of C6 glioma cells in a time-and dose-dependent manner. The mechanism of tamoxifen suppressing C6 glioma cells may be inhibiting proliferation and inducing apoptosis. Therefore, tamoxifen can be a candidate as a chemotherapy agent for glioma.展开更多
The 45, 55, 65 and 100 kDa ATP-binding proteinases (ATP-BPases) of the heat-shocked (44 ℃ for 30 min, recovery for 12h) rat C6 glioma cells were purified by DEAE-ionexchange and ATP-affinity chromatography. Their mol...The 45, 55, 65 and 100 kDa ATP-binding proteinases (ATP-BPases) of the heat-shocked (44 ℃ for 30 min, recovery for 12h) rat C6 glioma cells were purified by DEAE-ionexchange and ATP-affinity chromatography. Their molecular masses, isoelectric points (pI), pH-optima and other properties were analyzed by native proteinase gels.It was shown that the 65 kDa ATP-BPase is specifically induced by heat shock and not detectable in control cells.Its N-terminal 1-9 amino acid sequence was determined by Edman degradation, but no homologies to other proteins in the protein data bases were found. 30 and 31 kDa proteinases can be cleaved from the 45, 55 and 65 kDa proteinases to which they are linked. A possible relationship of the heat-induced 65 kDa ATP-BPase with the ATP-dependent proteinases (ATP-DPases) in prokaryotes and eukaryotes is discussed.展开更多
Using whole-cell patch-clamp recordings, the effects of antigliomatin were observed on chloride channels on C6 glioma cells cultured in vitro. Antigliomatin was extracted from the venom of the scorpion Buthus martensi...Using whole-cell patch-clamp recordings, the effects of antigliomatin were observed on chloride channels on C6 glioma cells cultured in vitro. Antigliomatin was extracted from the venom of the scorpion Buthus martensii Karsch. Chloride channels are closed under normal osmotic pressure. When osmotic pressure was reduced to 120, 110 and 100 mV, the cell volume enlarged, chloride channels opened, and the chloride channel current increased. Three minutes after antigliomatin treatment, the chloride channel current decreased in a dose-dependent manner. These results show that antigliomatin extracted from the venom of the scorpion Buthus martensii Karsch diminishes chloride channel currents on C6 glioma cells.展开更多
BACKGROUND:Studies have shown that cyclooxygenase-2 is associated with proliferation and apoptosis of glioma cells. OBJECTIVE:To investigate the effects of selective cyclooxygenase-2 inhibitor celecoxib on proliferati...BACKGROUND:Studies have shown that cyclooxygenase-2 is associated with proliferation and apoptosis of glioma cells. OBJECTIVE:To investigate the effects of selective cyclooxygenase-2 inhibitor celecoxib on proliferation and apoptosis of C6 glioma cells in vitro. DESIGN,TIME AND SETTING:A cellular,molecular,controlled study was performed at the Central Laboratory and Room of Electron Microscope,Medical School,Xi'an Jiaotong University,China from March 2007 to March 2008. MATERIALS:C6 glioma cells during in vitro log phase were assigned to control and experimental groups.Celecoxib(Pfizer,USA),dimethyl sulfoxide(Sigma,USA),and MTT(Sigma,USA) were used for this study. METHODS:The control group was subdivided into blank control and dimethyl sulfoxide control groups.C6 glioma cells in the blank control and dimethyl sulfoxide control groups were incubated in Dulbecco's modified Eagle's medium supplemented with 10%calf serum and 0.3%dimethyl sulfoxide respectively.C6 glioma cells in the experimental group were separately treated with 60,80, and 100μmol/L celecoxib. MAIN OUTCOME MEASURES:Activity of C6 glioma cells was examined by MTT assay.C6 glioma cell cycle and apoptosis were determined by annexin V-fluorescein isothiocyanate/propidium iodide double-staining,followed by flow cytometry.Morphology and ultrastructure of C6 glioma cells were observed with an inverted microscope and a transmission electron microscope,respectively. RESULTS:Compared with the blank control group,cell density was reduced,adherence ability weakened,and irregular nuclei were visible,with the presence of chromatin condensation, margination,and some apoptotic bodies in the experimental group.Activity of C6 glioma cells was significantly decreased(P<0.05),cell number was reduced during S phase,cell number was significantly increased during G_2/M phase(P<0.01),and the apoptotic rate was significantly increased(P<0.05) in the experimental group.These results were displayed in a dose- and time-dependent fashion.The outcomes were obvious in the 100μmol/L celecoxib group following 72 hours of treatment. CONCLUSION:Celecoxib blocked proliferation and induced apoptosis of C6 glioma cells in a dose-and time-dependent fashion.展开更多
BACKGROUND: Pseudomonas aeruginosa mannose-sensitive hemagglutinin (PA-MSHA) parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor ce...BACKGROUND: Pseudomonas aeruginosa mannose-sensitive hemagglutinin (PA-MSHA) parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor cell growth. OBJECTIVE: To investigate inhibitory mechanisms of PA-MSHA-induced proliferation in rat C6 glioma cells in vitro. DESIGN, TIME AND SETTING: Comparative observation and in vitro experiments were performed at the Key Laboratory of Natural Medicine, Kunming Medical College, China from July 2008 to April 2009. MATERIALS: Rat C6 glioma cell line (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China) and PA-MSHA parenteral injection (Beijing Wanteer Bio-Pharmaceutical, China) were used in the present study. METHODS: Rat C6 glioma cells in logarithmic growth phase were harvested in vitro. Adherent monolayer cells were respectively treated with PA-MSHA at final colony-forming units (cfu) of 1 × 108 cfu/mL, 2 × 108 cfu/mL, 4 × 108 cfu/mL, 6 × 108 cfu/mL, and 8 × 108 cfu/mL following 24 hours of conventional culture. MAIN OUTCOME MEASURES: MTT colorimetric assay was utilized to determine the inhibitory rate of C6 glioma cells following treatment with various concentrations of PA-MSHA at different times. Cell apoptosis was detected by fluorescent microscopy following Hoechst 33258 staining. Flow cytometry was used to measure PA-MSHA effects on C6 cell cycle. RESULTS: Inhibitory rate of C6 glioma cells increased with prolonged time and increased dose. Hoechst 33258 staining revealed obvious morphological changes in apoptotic C6 glioma cells. Flow cytometry revealed hypodiploid peaks, i.e., apoptotic peak, and the apoptotic rate in cells during S-phase significantly increased with increased concentrations in the experimental groups. CONCLUSION: With in vitro experiments, PA-MSHA preparations inhibited C6 glioma cell proliferation in a time- and dose-dependent manner. These mechanisms are likely associated with cell apoptosis induction and inhibition of the S phase.展开更多
Objective: To study the role of connexin gene (Cx 43) on the development of glioma and the feasibility of using Cx43cDNA as a target of gene therapy of gliomas. Methods: Parental rat C6 cells and C6 cells transfected ...Objective: To study the role of connexin gene (Cx 43) on the development of glioma and the feasibility of using Cx43cDNA as a target of gene therapy of gliomas. Methods: Parental rat C6 cells and C6 cells transfected with Cx43cDNA were implanted into right caudate nucleus of SD rats as control and transfected group. Rats bearing cerebral C6 gliomas were treated with Cx43cDNA and empty vector as treated group and empty vector group. The general manifestation, survival time, MRI dynamic scanning and histopathological changes of all rats were observed. In situ hybridization and immunohisto- chemistry were used for examination of Cx43mRNA and its protein in gliomas. Average number of AgNOR staining was used for detection of cell proliferation activity, and TUNEL method for determination of cell apoptosis. Results: All rats in control and empty vector group died of cerebral gliomas within 3 weeks after implantation of C6 cells. Six out of nine rats in the transfected group and eight out of ten rats in treated group kept alive beyond 120 days with totally disappearing of the tumor foci, except one treated rat having a little residue of tumor. In gliomas of transfected and treated groups Cx43 gene expression was upregulated, proliferation activity was lowered, However, the apoptotic cells did not increase. Conclusion: The present study indicates that Cx43 gene is of crucial importance in the development of malignant glioma. It can be an effective target for gene therapy of gliomas.展开更多
BACKGROUND:Human gliomas are more likely to express basic fibroblast growth factor-2(FGF-2), insulin-like growth factor-1(IGF-1),and IGF-1 receptor(IGF-1R) than normal brain tissue.These factors activate signal transd...BACKGROUND:Human gliomas are more likely to express basic fibroblast growth factor-2(FGF-2), insulin-like growth factor-1(IGF-1),and IGF-1 receptor(IGF-1R) than normal brain tissue.These factors activate signal transduction systems of Ras/MAPK and PI3K/Akt,which promote glioma growth. OBJECTIVE:To utilize RNA interference(RNAi) technique to down-regulate FGF-2,IGF-1,and IGF-1R gene expression,and to investigate the effects of these genes on rat C6 glioma cells,as well as the feasibility of RNAi for treating glioma. DESIGN,TIME AND SETTING:This neurooncological,randomized,controlled,in vivo and in vitro experiment,which used RNAi methodology,was performed at the Laboratory of Molecular Biology, Institute of Biochemistry,Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS:Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences.Small interfering RNA(siRNA) was synthesized by Shanghai GenePharma.Anti-IGF-1,anti-IGF-1R,anti-FGF-2,anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology,USA.Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center,Chinese Academy of Sciences. METHODS:C6 glioma cells were transfected with siRNA,which was chemically synthesized in vitro to correspond to endogenous FGF-2,IGF-1,and IGF-1R genes.The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR,and protein expression was determined by Western blot analysis.C6 glioma cell proliferation was observed using a growth curve. C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry.C6 glioma cell growth regression was observed by transwell migration assay.In addition,nude mouse subcutaneous tumor models were used in this study.For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA,two blank control groups,with six mice each,were set up:A(2.5μg siRNA was injected one week after C6 cells were inoculated,i.e.,when tumor volume reached 8 mm×8 mm) and B(siRNA was injected at the same time with C6 cells were inoculated.To study the effects of FGF-2 siRNA, the groups consisted of a blank control group,negative control group,2.6μg siRNA group,4μg siRNA group,and 5.3μg siRNA group,with six mice each. MAIN OUTCOME MEASURES:mRNA and protein inhibition ratio of FGF-2,IGF-1,and IGF-1R;C6 glioma cell proliferation,apoptosis,and cycle growth arrest;C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS:All siRNA constructs proved to be effective.After 48 hours,transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio>80%and an IGF-1 gene inhibition ratio of approximately 70%.Protein expression levels for FGF-2,IGF-1,and IGF-1R decreased in a dose-dependent manner following siRNA transfection,with an inhibition rate>85%,60%,and 50%, respectively.C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA.The apoptosis rate of C6 glioma cells induced by FGF-2,IGF-1,and IGF-1R siRNA was 39.96%,15.07%, and 22.47%,respectively(P<0.01).Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration.At 30 days after intratumoral injection of 2.6,4,and 5.3μg FGF-2 siRNA,tumor growth regression rate of FGF-2 siRNA was 56%,67%,and 86%,respectively. The tumor growth regression rate was 71.88%and 45.71%,respectively,when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation.When IGF-1 or IGF-1R siRNA was intratumorally injected during C6 glioma cell transplantation,the tumor growth regression rate was 78.13%and 74.29%,respectively. CONCLUSION:siRNA transfection downregulated gene expression of FGF-2,IGF-1,and IGF-1R. In addition,siRNA treatment markedly suppressed glioma cell proliferation,growth,and migration, and concomitantly reduced subcutaneous tumorigenicity.展开更多
Hydroxycamptothecin is a potent antineoplastic agent that has shown efficacy against multiple tumor lines in vitro. This is the first study to investigate the release, distribution, and efficacy of hydroxycamptothecin...Hydroxycamptothecin is a potent antineoplastic agent that has shown efficacy against multiple tumor lines in vitro. This is the first study to investigate the release, distribution, and efficacy of hydroxycamptothecin which was incorporated into the biodegradable polymer Polylactic Acid (PLA), and implant into brain directly. In vitro release curve generated showed that a large initial release occurred over the first three days and was followed by a steady, but considerably slower rate of release over the next 25 days. After implanting the discs into 40 male SD rats, the animals were followed up to 28 days, where the concentration in brain tissue was far higher than that in peripheral blood at the each of the eight time-points evaluated, and it was also within the therapeutic range for C6 cells tested in vitro. The in vivoefficacy of the discs was evaluated with rats inoculated intracranially with C6 glioma and treated with hydroxycamptothecin polymer compared to intravenous as well as intratumoral injections;the median survival is 21.1, 13.9, 14.9 days, respectively. Given these data, we conclude that the biodegradable polymer PLA releases hydroxycamptothecin, producing tumoricidal levels in brain tissues and prolonging survival in a rat glioma model.展开更多
Ganglioside and sulfatide are primary components of acidic glycosphingolipids(AGSLs),which are abundant in the brain tissue.AGSLs are potential tumor markers.In this paper,we use ultra-high performance liquid chromato...Ganglioside and sulfatide are primary components of acidic glycosphingolipids(AGSLs),which are abundant in the brain tissue.AGSLs are potential tumor markers.In this paper,we use ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to generate a first-ever comprehensive profile of AGSLs in brain and plasma of C6 glioma rats treated with temozolomide(TMZ).The AGSLs detected consisted mainly of C18-/C20-sphingosine.12,20,19,14 and 12 species were identified in GM1,GD1a,GD1b,GM3 and GT1b ganglioside groups which were abundant in rat brain,respectively.These five groups accounted for 88-89%of total ganglioside content.However,no AGSLs were detected in rat plasma.Possible biomarkers for abnormal changes in the glioma model and the protective effect of TMZ were mainly found in these ganglioside groups and sulfatide.The main lipid component of central and peripheral nervous system myelin sheathes is sulfatide,which is upregulated in many tumors.Antitumor properties of TMZ are due to modulation of sulfatide levels in tumor tissues.展开更多
Objectives: To elucidate how ethanol extract of L. serratum(ELS) could exert anti-migratory effects on glioma with the suppression of nuclear factor kappa B(NF-κB) downstream pathway. Methods: Cell viability of ELS o...Objectives: To elucidate how ethanol extract of L. serratum(ELS) could exert anti-migratory effects on glioma with the suppression of nuclear factor kappa B(NF-κB) downstream pathway. Methods: Cell viability of ELS on C6 glioma was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. Nitric oxide(NO) assay and 2',7'-dichlorofluorescin diacetate(DCFH-DA) assay were applied to measure NO production and reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced C6 glioma cells. NF-κB, mitogen-activated protein kinase(MAPK), inducible nictric oxide synthase(i NOS) and cyclooxygenase-2(COX-2) protein were determined by Western blot. Wound healing assay was used to investigate the inhibitory effect of ELS on fetal bovine serum(FBS)-induced migration and matrix metalloproteinase(MMP)-9 and-2 activity was examined by zymography. Results: ELS suppressed LPS-induced phosphorylation of extracellular signal-regulated kinase(ERK), c-Jun N-terminal kinase(JNK), and p38 through inhibiting the expression of chemokine CCL2(or monocyte chemoattractant protein-1, MCP-1). In addition, ELS inhibited the expression of i NOS, COX-2, and the production of NO by LPS in C6 glioma cells. ELS also significantly decreased serum-induced migration of C6 glioma cells in scratch wound healing in a dose-dependent manner(P<0.01). The activity of MMP-9 and-2 were also significantly attenuated by ELS with LPS treatment(P<0.01). Conclusion: Our results suggest that downregulation of MMP-9 gene expression might be involved in the anti-migration effect of ELS against LPS-induced C6 glioma cells.展开更多
Background: This paper describes the establishment of a rat intramedullary spinal cord tumor (IMSCT) model and histopathological characterization of the tumor model. Methods: Fourteen male Wistar rats were randomized ...Background: This paper describes the establishment of a rat intramedullary spinal cord tumor (IMSCT) model and histopathological characterization of the tumor model. Methods: Fourteen male Wistar rats were randomized into two groups. The rats in group 1 (control group, n = 7) received a 5 μl intramedullary injection of serum physiologic (SF). Those in group 2 (experimental group, n= 7) received a 5 μl intramedullary implantation of media containing 5 × 105 C6 glioma cells. The animals were sacrificed for histopathological examination at 21 days. Results: The control group showed normal functional and histopathological findings. The group 2 rats implanted with C6 glioblastoma cells developed hind-limb paraplegia. Pathological sections confirmed intramedullary C6 glioblastoma invading the spinal cord. Conclusions: A rat C6 IMSCT model was successfully established. This model may be useful in increasing understanding of intramedullary spinal cord gliomas in humans.展开更多
文摘OBJECTIVE The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS First,the model of C6 glioma in Wistar rats was established and verified by hemotoxylin and eosin staining,immunohistochemical staining and magnetic resonance imaging(MRI).Then different doses of lapachol were gavaged and tumor volumes of the C6 glioma were detected by MRI.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phen-azinemethosulfate(PMS)assay,Hoechst33358 staining,AnnexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomeraseⅠ(TOPⅠ)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322 DNA relaxation assay.Molecular docking was used to predict the interaction of lapachol-TOPⅠand lapachol-TOPⅡ.TOP I and TOPⅡexpression levels in C6 cells were determined by Enzymelinked immunosorbent assay kits and real-time polymerase chain reaction(RT-PCR).RESULTS The rat C6 glioma model was successfully established.High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).MTS/PMS assay,Hoechst 33258 staining,AnnexinⅤ-FITC/PI staining,and comet assay showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cells in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠandⅡ.Molecular docking showed that lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡ.Enzyme-linked immunosorbent assay and RT-PCR showed that lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠand TOPⅡactivities,as wel as TOPⅡexpression.
文摘Successful chemotherapy with paclitaxel(PTX)is impeded by multidrug resistance(MDR)in tumor cells.In this study,lipid-albumin nanoassemblies co-loaded with borneol and paclitaxel(BOR/PTX LANs)were prepared to circumvent MDR in C6 glioma cells.The physiochemical properties including particle size,encapsulation efficiency and morphology were evaluated in vitro.Quantitative and qualitative investigations of cellular uptake were carried out in C6 glioma cells.The cytotoxicity of the BOR/PTX LANs was determined by MTT assay.After that,the tumor targeting was also evaluated in C6 glioma bearing mice by in vivo imaging analysis.BOR/PTX LANs have a higher entrapment efficiency(90.4±1.2%),small particle size(107.5±3.2 nm),narrow distribution(P.I.=0.171±0.02).The cellular uptake of PTX was significantly increased by BOR/PTX LANs compared with paclitaxel loaded lipidalbumin nanoassemblies(PTX LANs)in quantitative research.The result was further confirmed by confocal laser scanning microscopy qualitatively.The cellular uptake was energy-,timeand concentration-dependent,and clathrin-and endosome/lysosome-associated pathways were involved.The BOR/PTX LANs displayed a higher cytotoxicity agaist C6 glioma cells in comparion with PTX LANs and Taxol.Moreover,the encapsulation of BOR in LANs obviously increased the accumulation of the drug in tumor tissues,demonstrating the tumor targeted ability of BOR/PTX LANs.These results indicated that BOR/PTX LANs could overcome MDR by combination of drug delivery systems and P-gp inhibition,and shown the potential for treatment of gliomas.
基金Hubei Provincial Education Department Foundation, No. Q20092405Hubei Provincial Science and Technology Agency Foundation, No. 2005AA301C28Hubei Provincial Health Department Foundation, No. QJX2005-15
文摘BACKGROUND: Embryonic neural stem cells (NSCs) have provided positive effects for the treatment of glioma. However, the source for embryonic NSCs remains limited and high amplification conditions are required. Bone marrow stromal cells (BMSCs) have been proposed for the treatment of glioma. OBJECTIVE: To investigate biological changes in NSCs and BMSCs following transplantation into rat models of glioma. DESIGN, TIME AND SETTING: A randomized, controlled, animal experiment was performed at the Embryonic Stem Cell Research Laboratory of Yunyang Medical College from February 2006 to August 2008. MATERIALS: The rat C6 glioma cell line was purchased from Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; mouse anti-bromodeoxyuridine (BrdU) monoclonal antibody and Cy3-labeled goat anti-mouse IgG antibody was purchased from Upstate, USA. METHODS: A total of 95 Sprague Dawley rats were randomly assigned to three groups: NSC (n = 35), transplanted with > 6 × 106 NSCs via left medial hind limb; BMSC (n = 35), transplanted with > 1 × 106 BMSCs via left medial hind limb; model group (n = 25), injected with the same volume of 0.1 mmol/L phosphate buffered saline. MAIN OUTCOME MEASURES: Gliomal growth and size were assessed by nuclear magnetic resonance, and glioma morphological features were observed following hematoxylin-eosin staining and BrdU immunohistochemistry 3 and 4 weeks following transplantation. RESULTS: The average survival of rats in the BMSC, NSC, and model groups was 4.03, 4.28, and 3.88 weeks. At 3 weeks, there was no significant difference in the average glioma diameter between the BMSC and model groups (P > 0.05). However, gliomal diameter was significantly decreased in the NSC group compared with the model group (P < 0.05). At 4 weeks, there was no statistical difference between the groups (P > 0.05). BrdU immunohistochemistry revealed that BMSCs and NSCs appeared to migrate to the gliomas. CONCLUSION: NSCs inhibited glioma cell growth and prolonged rat survival. BMSCs did not significantly suppress glioma cell growth.
文摘OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities.The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo,as well as the potential mechanisms.METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats.The effects of lapachol on C6 cell proliferation,apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/phenazinemethosulfate(PMS)assay,hoechst 33358 staining,annexinⅤ-FITC/PI staining,and comet assay.Effects of lapachol on topoisomerase I(TOP I)and topoisomeraseⅡ(TOPⅡ)activities were detected by TOPⅠand TOPⅡmediated supercoiled p BR322DNA relaxation assays and molecular docking.TOPⅠand TOPⅡexpression levels in C6 cells were also determined.RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats(P<0.05).It was showed that lapachol could inhibit proliferation,induce apoptosis and DNA damage of C6 cel s in dose dependent manners.Lapachol could inhibit the activities of both TOPⅠ and Ⅱ.Lapachol-TOPⅠshowed relatively stronger interaction than that of lapachol-TOPⅡin molecular docking study.Also,lapachol could inhibit TOPⅡexpression levels,but not TOPⅠexpression levels.CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro,which might be related with inhibiting TOPⅠ and TOPⅡ activities,as wel as TOPⅡ expression.
文摘Objective To investigate the anti-tumor effect and mechanism of tamoxifen on rat C6 glioma cells. Methods C6 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 3% fetal calf serum (FCS), and treated with tamoxifen of different concentrations, i.e. group A (1.25μmol/L), group B (2.50 μmol/L), group C (5.00 μmol/L), group D (10.00 μmol/L), group E (20.00 μmol/L) and control group (0.00 μmol/L). Morphological changes, MTT assay and 5-bromo-2'-deoxyuriding labeling ratio were assessed. Apoptosis was observed by flow cytometry. Results C6 cells treated with different doses of tamoxifen for 24, 48, and 72 hours became irregular in shape, while cells treated with vehicle grew normally. MTT assay showed that tamoxifen did not suppress C6 cell growth until 72 hours after treatment. Seventy-two hours after treatment, there were significant differences in cell viable rate between group A versus groups C, D and E; so did group B versus group D as well as group E (P<0.05). BrdU incorporation assay indicated significant difference of BrdU labbled index (BrdU LI) among groups A, C, E and control group 48 hours after treatment (P<0.05). And the BrdU LI decreased with the increased concentration of tamoxifen. Flow cytometry (FCM) showed significant difference between treated group and control group at 24, 48, and 72 hours after treatment (P<0.05). Conclusion Tamoxifen significantly suppresses the growth of C6 glioma cells in a time-and dose-dependent manner. The mechanism of tamoxifen suppressing C6 glioma cells may be inhibiting proliferation and inducing apoptosis. Therefore, tamoxifen can be a candidate as a chemotherapy agent for glioma.
文摘The 45, 55, 65 and 100 kDa ATP-binding proteinases (ATP-BPases) of the heat-shocked (44 ℃ for 30 min, recovery for 12h) rat C6 glioma cells were purified by DEAE-ionexchange and ATP-affinity chromatography. Their molecular masses, isoelectric points (pI), pH-optima and other properties were analyzed by native proteinase gels.It was shown that the 65 kDa ATP-BPase is specifically induced by heat shock and not detectable in control cells.Its N-terminal 1-9 amino acid sequence was determined by Edman degradation, but no homologies to other proteins in the protein data bases were found. 30 and 31 kDa proteinases can be cleaved from the 45, 55 and 65 kDa proteinases to which they are linked. A possible relationship of the heat-induced 65 kDa ATP-BPase with the ATP-dependent proteinases (ATP-DPases) in prokaryotes and eukaryotes is discussed.
基金the Science and Technology Development Program of Jilin Province, No.20050407-6
文摘Using whole-cell patch-clamp recordings, the effects of antigliomatin were observed on chloride channels on C6 glioma cells cultured in vitro. Antigliomatin was extracted from the venom of the scorpion Buthus martensii Karsch. Chloride channels are closed under normal osmotic pressure. When osmotic pressure was reduced to 120, 110 and 100 mV, the cell volume enlarged, chloride channels opened, and the chloride channel current increased. Three minutes after antigliomatin treatment, the chloride channel current decreased in a dose-dependent manner. These results show that antigliomatin extracted from the venom of the scorpion Buthus martensii Karsch diminishes chloride channel currents on C6 glioma cells.
基金the National Natural Science Foundation of China,No.30672162
文摘BACKGROUND:Studies have shown that cyclooxygenase-2 is associated with proliferation and apoptosis of glioma cells. OBJECTIVE:To investigate the effects of selective cyclooxygenase-2 inhibitor celecoxib on proliferation and apoptosis of C6 glioma cells in vitro. DESIGN,TIME AND SETTING:A cellular,molecular,controlled study was performed at the Central Laboratory and Room of Electron Microscope,Medical School,Xi'an Jiaotong University,China from March 2007 to March 2008. MATERIALS:C6 glioma cells during in vitro log phase were assigned to control and experimental groups.Celecoxib(Pfizer,USA),dimethyl sulfoxide(Sigma,USA),and MTT(Sigma,USA) were used for this study. METHODS:The control group was subdivided into blank control and dimethyl sulfoxide control groups.C6 glioma cells in the blank control and dimethyl sulfoxide control groups were incubated in Dulbecco's modified Eagle's medium supplemented with 10%calf serum and 0.3%dimethyl sulfoxide respectively.C6 glioma cells in the experimental group were separately treated with 60,80, and 100μmol/L celecoxib. MAIN OUTCOME MEASURES:Activity of C6 glioma cells was examined by MTT assay.C6 glioma cell cycle and apoptosis were determined by annexin V-fluorescein isothiocyanate/propidium iodide double-staining,followed by flow cytometry.Morphology and ultrastructure of C6 glioma cells were observed with an inverted microscope and a transmission electron microscope,respectively. RESULTS:Compared with the blank control group,cell density was reduced,adherence ability weakened,and irregular nuclei were visible,with the presence of chromatin condensation, margination,and some apoptotic bodies in the experimental group.Activity of C6 glioma cells was significantly decreased(P<0.05),cell number was reduced during S phase,cell number was significantly increased during G_2/M phase(P<0.01),and the apoptotic rate was significantly increased(P<0.05) in the experimental group.These results were displayed in a dose- and time-dependent fashion.The outcomes were obvious in the 100μmol/L celecoxib group following 72 hours of treatment. CONCLUSION:Celecoxib blocked proliferation and induced apoptosis of C6 glioma cells in a dose-and time-dependent fashion.
文摘BACKGROUND: Pseudomonas aeruginosa mannose-sensitive hemagglutinin (PA-MSHA) parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor cell growth. OBJECTIVE: To investigate inhibitory mechanisms of PA-MSHA-induced proliferation in rat C6 glioma cells in vitro. DESIGN, TIME AND SETTING: Comparative observation and in vitro experiments were performed at the Key Laboratory of Natural Medicine, Kunming Medical College, China from July 2008 to April 2009. MATERIALS: Rat C6 glioma cell line (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China) and PA-MSHA parenteral injection (Beijing Wanteer Bio-Pharmaceutical, China) were used in the present study. METHODS: Rat C6 glioma cells in logarithmic growth phase were harvested in vitro. Adherent monolayer cells were respectively treated with PA-MSHA at final colony-forming units (cfu) of 1 × 108 cfu/mL, 2 × 108 cfu/mL, 4 × 108 cfu/mL, 6 × 108 cfu/mL, and 8 × 108 cfu/mL following 24 hours of conventional culture. MAIN OUTCOME MEASURES: MTT colorimetric assay was utilized to determine the inhibitory rate of C6 glioma cells following treatment with various concentrations of PA-MSHA at different times. Cell apoptosis was detected by fluorescent microscopy following Hoechst 33258 staining. Flow cytometry was used to measure PA-MSHA effects on C6 cell cycle. RESULTS: Inhibitory rate of C6 glioma cells increased with prolonged time and increased dose. Hoechst 33258 staining revealed obvious morphological changes in apoptotic C6 glioma cells. Flow cytometry revealed hypodiploid peaks, i.e., apoptotic peak, and the apoptotic rate in cells during S-phase significantly increased with increased concentrations in the experimental groups. CONCLUSION: With in vitro experiments, PA-MSHA preparations inhibited C6 glioma cell proliferation in a time- and dose-dependent manner. These mechanisms are likely associated with cell apoptosis induction and inhibition of the S phase.
基金This work was supported by the National Natural Science Foundation of China (No. 39870815).
文摘Objective: To study the role of connexin gene (Cx 43) on the development of glioma and the feasibility of using Cx43cDNA as a target of gene therapy of gliomas. Methods: Parental rat C6 cells and C6 cells transfected with Cx43cDNA were implanted into right caudate nucleus of SD rats as control and transfected group. Rats bearing cerebral C6 gliomas were treated with Cx43cDNA and empty vector as treated group and empty vector group. The general manifestation, survival time, MRI dynamic scanning and histopathological changes of all rats were observed. In situ hybridization and immunohisto- chemistry were used for examination of Cx43mRNA and its protein in gliomas. Average number of AgNOR staining was used for detection of cell proliferation activity, and TUNEL method for determination of cell apoptosis. Results: All rats in control and empty vector group died of cerebral gliomas within 3 weeks after implantation of C6 cells. Six out of nine rats in the transfected group and eight out of ten rats in treated group kept alive beyond 120 days with totally disappearing of the tumor foci, except one treated rat having a little residue of tumor. In gliomas of transfected and treated groups Cx43 gene expression was upregulated, proliferation activity was lowered, However, the apoptotic cells did not increase. Conclusion: The present study indicates that Cx43 gene is of crucial importance in the development of malignant glioma. It can be an effective target for gene therapy of gliomas.
基金the National Natural Science Foundation of China,No.30371459Science and Technology Development Fund of Shanghai,No.034047
文摘BACKGROUND:Human gliomas are more likely to express basic fibroblast growth factor-2(FGF-2), insulin-like growth factor-1(IGF-1),and IGF-1 receptor(IGF-1R) than normal brain tissue.These factors activate signal transduction systems of Ras/MAPK and PI3K/Akt,which promote glioma growth. OBJECTIVE:To utilize RNA interference(RNAi) technique to down-regulate FGF-2,IGF-1,and IGF-1R gene expression,and to investigate the effects of these genes on rat C6 glioma cells,as well as the feasibility of RNAi for treating glioma. DESIGN,TIME AND SETTING:This neurooncological,randomized,controlled,in vivo and in vitro experiment,which used RNAi methodology,was performed at the Laboratory of Molecular Biology, Institute of Biochemistry,Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS:Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences.Small interfering RNA(siRNA) was synthesized by Shanghai GenePharma.Anti-IGF-1,anti-IGF-1R,anti-FGF-2,anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology,USA.Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center,Chinese Academy of Sciences. METHODS:C6 glioma cells were transfected with siRNA,which was chemically synthesized in vitro to correspond to endogenous FGF-2,IGF-1,and IGF-1R genes.The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR,and protein expression was determined by Western blot analysis.C6 glioma cell proliferation was observed using a growth curve. C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry.C6 glioma cell growth regression was observed by transwell migration assay.In addition,nude mouse subcutaneous tumor models were used in this study.For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA,two blank control groups,with six mice each,were set up:A(2.5μg siRNA was injected one week after C6 cells were inoculated,i.e.,when tumor volume reached 8 mm×8 mm) and B(siRNA was injected at the same time with C6 cells were inoculated.To study the effects of FGF-2 siRNA, the groups consisted of a blank control group,negative control group,2.6μg siRNA group,4μg siRNA group,and 5.3μg siRNA group,with six mice each. MAIN OUTCOME MEASURES:mRNA and protein inhibition ratio of FGF-2,IGF-1,and IGF-1R;C6 glioma cell proliferation,apoptosis,and cycle growth arrest;C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS:All siRNA constructs proved to be effective.After 48 hours,transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio>80%and an IGF-1 gene inhibition ratio of approximately 70%.Protein expression levels for FGF-2,IGF-1,and IGF-1R decreased in a dose-dependent manner following siRNA transfection,with an inhibition rate>85%,60%,and 50%, respectively.C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA.The apoptosis rate of C6 glioma cells induced by FGF-2,IGF-1,and IGF-1R siRNA was 39.96%,15.07%, and 22.47%,respectively(P<0.01).Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration.At 30 days after intratumoral injection of 2.6,4,and 5.3μg FGF-2 siRNA,tumor growth regression rate of FGF-2 siRNA was 56%,67%,and 86%,respectively. The tumor growth regression rate was 71.88%and 45.71%,respectively,when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation.When IGF-1 or IGF-1R siRNA was intratumorally injected during C6 glioma cell transplantation,the tumor growth regression rate was 78.13%and 74.29%,respectively. CONCLUSION:siRNA transfection downregulated gene expression of FGF-2,IGF-1,and IGF-1R. In addition,siRNA treatment markedly suppressed glioma cell proliferation,growth,and migration, and concomitantly reduced subcutaneous tumorigenicity.
文摘Hydroxycamptothecin is a potent antineoplastic agent that has shown efficacy against multiple tumor lines in vitro. This is the first study to investigate the release, distribution, and efficacy of hydroxycamptothecin which was incorporated into the biodegradable polymer Polylactic Acid (PLA), and implant into brain directly. In vitro release curve generated showed that a large initial release occurred over the first three days and was followed by a steady, but considerably slower rate of release over the next 25 days. After implanting the discs into 40 male SD rats, the animals were followed up to 28 days, where the concentration in brain tissue was far higher than that in peripheral blood at the each of the eight time-points evaluated, and it was also within the therapeutic range for C6 cells tested in vitro. The in vivoefficacy of the discs was evaluated with rats inoculated intracranially with C6 glioma and treated with hydroxycamptothecin polymer compared to intravenous as well as intratumoral injections;the median survival is 21.1, 13.9, 14.9 days, respectively. Given these data, we conclude that the biodegradable polymer PLA releases hydroxycamptothecin, producing tumoricidal levels in brain tissues and prolonging survival in a rat glioma model.
基金The Ministry of Science and Technology of the People’s Republic of China[2016ZX09101017]supported this project.
文摘Ganglioside and sulfatide are primary components of acidic glycosphingolipids(AGSLs),which are abundant in the brain tissue.AGSLs are potential tumor markers.In this paper,we use ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry to generate a first-ever comprehensive profile of AGSLs in brain and plasma of C6 glioma rats treated with temozolomide(TMZ).The AGSLs detected consisted mainly of C18-/C20-sphingosine.12,20,19,14 and 12 species were identified in GM1,GD1a,GD1b,GM3 and GT1b ganglioside groups which were abundant in rat brain,respectively.These five groups accounted for 88-89%of total ganglioside content.However,no AGSLs were detected in rat plasma.Possible biomarkers for abnormal changes in the glioma model and the protective effect of TMZ were mainly found in these ganglioside groups and sulfatide.The main lipid component of central and peripheral nervous system myelin sheathes is sulfatide,which is upregulated in many tumors.Antitumor properties of TMZ are due to modulation of sulfatide levels in tumor tissues.
基金Supported by Dongguk University Research Fund of 2015
文摘Objectives: To elucidate how ethanol extract of L. serratum(ELS) could exert anti-migratory effects on glioma with the suppression of nuclear factor kappa B(NF-κB) downstream pathway. Methods: Cell viability of ELS on C6 glioma was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. Nitric oxide(NO) assay and 2',7'-dichlorofluorescin diacetate(DCFH-DA) assay were applied to measure NO production and reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced C6 glioma cells. NF-κB, mitogen-activated protein kinase(MAPK), inducible nictric oxide synthase(i NOS) and cyclooxygenase-2(COX-2) protein were determined by Western blot. Wound healing assay was used to investigate the inhibitory effect of ELS on fetal bovine serum(FBS)-induced migration and matrix metalloproteinase(MMP)-9 and-2 activity was examined by zymography. Results: ELS suppressed LPS-induced phosphorylation of extracellular signal-regulated kinase(ERK), c-Jun N-terminal kinase(JNK), and p38 through inhibiting the expression of chemokine CCL2(or monocyte chemoattractant protein-1, MCP-1). In addition, ELS inhibited the expression of i NOS, COX-2, and the production of NO by LPS in C6 glioma cells. ELS also significantly decreased serum-induced migration of C6 glioma cells in scratch wound healing in a dose-dependent manner(P<0.01). The activity of MMP-9 and-2 were also significantly attenuated by ELS with LPS treatment(P<0.01). Conclusion: Our results suggest that downregulation of MMP-9 gene expression might be involved in the anti-migration effect of ELS against LPS-induced C6 glioma cells.
文摘Background: This paper describes the establishment of a rat intramedullary spinal cord tumor (IMSCT) model and histopathological characterization of the tumor model. Methods: Fourteen male Wistar rats were randomized into two groups. The rats in group 1 (control group, n = 7) received a 5 μl intramedullary injection of serum physiologic (SF). Those in group 2 (experimental group, n= 7) received a 5 μl intramedullary implantation of media containing 5 × 105 C6 glioma cells. The animals were sacrificed for histopathological examination at 21 days. Results: The control group showed normal functional and histopathological findings. The group 2 rats implanted with C6 glioblastoma cells developed hind-limb paraplegia. Pathological sections confirmed intramedullary C6 glioblastoma invading the spinal cord. Conclusions: A rat C6 IMSCT model was successfully established. This model may be useful in increasing understanding of intramedullary spinal cord gliomas in humans.
