)| PMID |
10671549 ( ) |
|---|---|
| Title | Cu,Zn-superoxide dismutase-dependent apoptosis induced by nitric oxide in neuronal cells. |
| Abstract | Nitric oxide (NO) challenge to human neuroblastoma cells (SH-SY5Y) ultimately results in apoptosis. Tumor suppressor protein p53 and cell cycle inhibitor p21 accumulate as an early sign of S-nitrosoglutathione-mediated toxicity. Cytochrome c release from mitochondria and caspase 3 activation also occurred. Cells transfected with either wild type (WT) or mutant (G93A) Cu, Zn-superoxide dismutase (Cu,Zn-SOD) produced comparable amounts of nitrite/nitrate but showed different degree of apoptosis. G93A cells were the most affected and WT cells the most protected; however, Cu, Zn-SOD content of these two cell lines was 2-fold the SH-SY5Y cells under both resting and treated conditions. We linked decreased susceptibility of the WT cells to higher and more stable Bcl-2 and decreased reactive oxygen species. Conversely, we linked G93A susceptibility to increased reactive oxygen species production since simultaneous administration of S-nitrosoglutathione and copper chelators protects from apoptosis. Furthermore, G93A cells showed a significant decrease of Bcl-2 expression and, as target of NO-derived radicals, showed lower cytochrome c oxidase activity. These results demonstrate that resistance to NO-mediated apoptosis is strictly related to the level and integrity of Cu,Zn-SOD and that the balance between reactive nitrogen and reactive oxygen species regulates neuroblastoma apoptosis. via dei Vestini, 66100 Chieti, Italy. Ciriolo@bio.uniroma2.it |
NOTE: Color highlight is limited to the abstract and SciMiner text-mining mode. If you see much more identified targets below from "Targets by SciMiner Summary" and "Targets by SciMiner Full list", they may have been identified from the full text.
Targets by SciMiner Summary
| HUGO ID | Symbol | Target Name | #Occur | ActualStr |
|---|---|---|---|---|
| 990 | BCL2 | B-cell CLL/lymphoma 2 | 45 | Bcl-2 | bcl2 | bcl 2 | Bcl2 | |
| 19986 | CYCS | cytochrome c, somatic | 24 | cytochrome c | |
| 11998 | TP53 | tumor protein p53 | 23 | p53-mediated | |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | 22 | SOD | superoxide dismutase | |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | 15 | p21 | |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | 13 | caspase 3 | |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | 7 | cytochrome c oxidase | |
| 11180 | SOD2 | superoxide dismutase 2, mitochondrial | 4 | mn sod | Mn-SOD | |
| 13716 | C4orf6 | chromosome 4 open reading frame 6 | 1 | expressed in neuroblastoma | |
| 5173 | HRAS | v-Ha-ras Harvey rat sarcoma viral oncogene homolog | 1 | p21 protein | |
| 992 | BCL2L1 | BCL2-like 1 | 1 | bcl xl | |
| 2295 | CP | ceruloplasmin (ferroxidase) | 1 | ceruloplasmin | |
| 5472 | IGFBP3 | insulin-like growth factor binding protein 3 | 1 | bp53 | |
| 3701 | FHIT | fragile histidine triad gene | 1 | tumor suppressor protein | |
Targets by SciMiner Full list
| HUGO ID | Symbol | Name | ActualStr | Score | FlankingText |
|---|---|---|---|---|---|
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Tumor suppressor protein p53 and cell cycle inhibitor p21 accumulate as an early sign |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | Tumor suppressor protein p53 and cell cycle inhibitor p21 accumulate as an early sign of S -nitrosoglutathione-mediated toxicity |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | susceptibility of the WT cells to higher and more stable Bcl-2 and decreased reactive oxygen species |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | Furthermore G93A cells showed a significant decrease of Bcl-2 expression and as target of NO-derived radicals showed lower cytochrome |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | Constitutive expression of high Bcl-2 protein levels by transfection experiments has proven that Bcl-2 or |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | high Bcl-2 protein levels by transfection experiments has proven that Bcl-2 or related family members can protect from NO-mediated apoptosis ( |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | The tumor suppressor gene product p53 is able to modulate apoptosis in DNA-damaged cell ( 17 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | The enzyme superoxide dismutase (SOD) SOD may play a fundamental role in modulating NO toxicity since |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | As matter of fact cells producing an increased amount of SOD were resistant to NO-mediated toxicity ( 24 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | sclerosis (FALS FALS are pro-apoptotic agents although they retain enzymatic SOD activity ( 26 |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | cells is associated with a canonical sequence of events including Bcl-2 p53 and p21 modulation cytochrome c release in the cytosol |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | is associated with a canonical sequence of events including Bcl-2 p53 and p21 modulation cytochrome c release in the cytosol and |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | with a canonical sequence of events including Bcl-2 p53 and p21 modulation cytochrome c release in the cytosol and caspase 3 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | Cells carrying wild type SOD are protected from NO-mediated apoptosis whereas cells transfected with the |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | protected from NO-mediated apoptosis whereas cells transfected with the mutant SOD are more susceptible |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | type Cu Zn-SOD (named named WT or with a mutated SOD G93A (named named G93A were obtained as described previously ( |
| 11180 | SOD2 | superoxide dismutase 2, mitochondrial | Mn-SOD | 1.9 | Transfected cells had the same antioxidant pattern such as of Mn-SOD as SH-SY5Y cells |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | p53 p21 and Bcl-2 Proteins Detection--Cell pellet was resuspended in lysis |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | p53 p21 and Bcl-2 Proteins Detection--Cell pellet was resuspended in lysis buffer |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | p53 p21 and Bcl-2 Proteins Detection--Cell pellet was resuspended in lysis buffer containing 10 |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | microg of proteins were loaded on a 10 (for for p53 or 12% (for for p21 and Bcl-2 polyacrylamide gel |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | on a 10 (for for p53 or 12% (for for p21 and Bcl-2 polyacrylamide gel |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | 10 (for for p53 or 12% (for for p21 and Bcl-2 polyacrylamide gel |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | Supernatants were used for SOD activity and protein content assay |
| 11180 | SOD2 | superoxide dismutase 2, mitochondrial | Mn-SOD | 1.9 | Under these conditions the activity of Mn-SOD is almost fully inhibited ( 33 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | However cells transfected with normal SOD were much less protected than against NO-mediated apoptosis |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | be strengthened by O 2 via formation of peroxynitrite and SOD that catalytically removes O 2 is therefore indicated as a |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | However cells transfected with a SOD mutant (G93A G93A as active as the wild type were |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | NO-mediated apoptosis and that additional properties of the G93A mutant SOD potentiate apoptogenic stimuli of NO |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | In our model apoptosis was characterized by an induction of p53 protein which is able to induce either growth arrest or |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | is dependent upon sequence-specific DNA binding and transcriptional activation of p53 target genes such as p21 which is an inhibitor of |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | binding and transcriptional activation of p53 target genes such as p21 which is an inhibitor of cyclin-dependent kinases and thereby blocks |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | cyclin-dependent kinases and thereby blocks cell cycle progression ( 42 p53 increase was an early response to GSNO treatment in fact |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | entered the apoptotic pathway they showed the same degree of p53 accumulation as the other cell lines tested |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Furthermore p53 accumulation and activation leads to p21 increase in all the |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | Furthermore p53 accumulation and activation leads to p21 increase in all the three cell types however the cleavage |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | all the three cell types however the cleavage product of p21 was observed only in the G93A and SH-SY5Y cells at |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | It has been demonstrated that p21 cleavage could be reproduced in extracts prepared from irradiated cells |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | From these observations it was suggested that p21 may serve as a critical checkpoint regulator for both cell |
| 11998 | TP53 | tumor protein p53 | p53-mediated | 0.6 | regulator for both cell cycle arrest and apoptosis during the p53-mediated DNA damage response ( 42 |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | on Bax and/or and or Bid two members of the Bcl-2 protein family ( 57 |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | In our experiments Bcl-2 was down-regulated by the GSNO treatment in SH-SY5Y cells whereas |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | prone to NO-induced apoptosis we propose a causative role for Bcl-2 in the increased susceptibility of G93A cells to apoptosis |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | It has been suggested that the antiapoptotic effects of Bcl-2 be at least in part due to its antioxidant activity |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | modulators of physiological signal transduction ( 60 is increased when Bcl-2 is down-regulated |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | a factor protecting neurons from NO-induced apoptosis whereas the G93A SOD mutant despite high dismutating activity increased intracellular flux of ROS |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | A vicious circle including down-regulation of Bcl-2 and deviating redox activity of copper in the G93A mutant |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Immunoreactive p53 protein ( C and immunoreactive p21 protein ( D were |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | Immunoreactive p53 protein ( C and immunoreactive p21 protein ( D were measured by Western blotting using monoclonal |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | Bcl-2 content of SH-SY5Y cells and effects on it by Cu |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | Bcl-2 immunoreactive protein was detected by Western blotting using a monoclonal |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | SOD activity (not not shown and protein levels of WT and |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | Furthermore SOD activity of the three cell lines was unaffected by GSNO |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | by GSNO treatment (not not shown as well as the SOD protein content as assessed by Western blot analysis (Fig Fig |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Apoptotic Markers Cytochrome c Release Caspase Activation p53 Accumulation p21 Increase and Bcl2 Down-regulation-- To examine the sequence |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | Apoptotic Markers Cytochrome c Release Caspase Activation p53 Accumulation p21 Increase and Bcl2 Down-regulation-- To examine the sequence of events |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl2 | 1.0 | Cytochrome c Release Caspase Activation p53 Accumulation p21 Increase and Bcl2 Down-regulation-- To examine the sequence of events occurring upon GSNO |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | The tumor suppressor gene p53 is known to be a member of the DNA damage-response |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | It has been demonstrated that p53 protein increases in response to NO donors and that it |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Fig 3 C shows that the p53 protein was stably expressed in neuroblastoma cells to a varying |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | a varying extent in particular WT cells showed a lower p53 protein level with respect to SH-SY5Y cells and G93A cells |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | Upon GSNO treatment p53 significantly accumulated even in the WT cells reaching comparable values |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | almost disappeared after GSNO treatment supporting the current opinion that p53 activation involves phosphorylation of the protein |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | In response to DNA damage human p53 is phosphorylated within its transactivation domain at serine 15 and |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | In order to confirm further a role for p53 activation in the GSNO-mediated toxicity we analyze the expression of |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | activation in the GSNO-mediated toxicity we analyze the expression of p21 which is a potent inhibitor of cell cycle kinases also |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | It has been demonstrated that ectopic expression of p53 alone could induce p21 expression followed by p21 cleavage and |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | been demonstrated that ectopic expression of p53 alone could induce p21 expression followed by p21 cleavage and apoptosis ( 43 |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | expression of p53 alone could induce p21 expression followed by p21 cleavage and apoptosis ( 43 |
| 1784 | CDKN1A | cyclin-dependent kinase inhibitor 1A (p21, Cip1) | p21 | 1.4 | Fig 3 D shows that p21 is expressed in SH-SY5Y cells and that the protein accumulated |
| 11998 | TP53 | tumor protein p53 | p53 | 5.8 | a 2-fold indication as follows first it is confirmatory of p53 activation and second it indicates that G93A (at at higher |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | Bcl-2 protein modulation is responsible for cell survival or suicide constitutive |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | responsible for cell survival or suicide constitutive expression of high Bcl-2 protein levels by transfection experiments has proven that Bcl-2 or |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | high Bcl-2 protein levels by transfection experiments has proven that Bcl-2 or related family members can protect cells from NO-mediated apoptosis |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | analysis untreated SH-SY5Y and WT cells express high levels of Bcl-2 protein whereas G93A cells show a much lower content (Fig |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | content (Fig Fig 4 48 h of GSNO treatment decreased Bcl-2 expression although to a different extent in particular Bcl-2 content |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | Bcl-2 | 4.3 | decreased Bcl-2 expression although to a different extent in particular Bcl-2 content of G93A cells was at the limit of detection |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | apoptosis of G93A cells cannot be explained in terms of SOD level |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | or hydroxyl radical formation has been demonstrated for fully active SOD mutants associated with FALS ( 47 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 2.4 | not significantly affected either by the transfection with the mutant SOD or by the GSNO treatment |
| 3701 | FHIT | fragile histidine triad gene | tumor suppressor protein | 1.0 | tumor suppressor protein p53 and cell cycle inhibitor p21 accumulate as an early sign of s nitrosoglutathione mediated toxicity. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | cytochrome c release from mitochondria and caspase 3 activation also occurred. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | cytochrome c release from mitochondria and caspase 3 activation also occurred. |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | cells transfected with either wild type wt or mutant g93a cu zn superoxide dismutase cu zn sod produced comparable amounts of nitrite/nitrate but showed different degree of apoptosis. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | we linked decreased susceptibility of the wt cells to higher and more stable bcl 2 and decreased reactive oxygen species. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | furthermore g93a cells showed a significant decrease of bcl 2 expression and as target of no derived radicals showed lower cytochrome c oxidase activity. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | furthermore g93a cells showed a significant decrease of bcl 2 expression and as target of no derived radicals showed lower cytochrome c oxidase activity. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | among these the protein product of bcl 2 protooncogene plays a pivotal role in apoptosis by regulating cytochrome c efflux from mitochondria which in turn activates cysteine proteases caspases ultimately leading to specific dna fragmentatio |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | among these the protein product of bcl 2 protooncogene plays a pivotal role in apoptosis by regulating cytochrome c efflux from mitochondria which in turn activates cysteine proteases caspases ultimately leading to specific dna fragmentation. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | constitutive expression of high bcl 2 protein levels by transfection experiments has proven that bcl 2 or related family members can protect from no mediated apoptosis 15 16 . |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | the enzyme superoxide dismutase sod may play a fundamental role in modulating no toxicity since it acts as an antioxidant dismutating o 2 . |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | the results obtained showed that gsno mediated apoptosis in these cells is associated with a canonical sequence of events including bcl 2 p53 and p21 modulation cytochrome c release in the cytosol and caspase 3 activation. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | he results obtained showed that gsno mediated apoptosis in these cells is associated with a canonical sequence of events including bcl 2 p53 and p21 modulation cytochrome c release in the cytosol and caspase 3 activation. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | the results obtained showed that gsno mediated apoptosis in these cells is associated with a canonical sequence of events including bcl 2 p53 and p21 modulation cytochrome c release in the cytosol and caspase 3 activation. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | se inhibitor mixture dithiothreitol dtt sulfanilamide p aminobenzenesulfonamide phenylmethylsulfonyl fluoride n 1 naphthyl ethylenediamine dihydrochloride bathocuproinedisulfonic acid monoclonal anti bcl 2 clone bcl 2 100 monoclonal anti p53 clone bp53 12 and cytochrome c horse heart were obtained from sigma. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | clone bcl 2 100 monoclonal anti p53 clone bp53 12 and cytochrome c horse heart were obtained from sigma. |
| 5472 | IGFBP3 | insulin-like growth factor binding protein 3 | bp53 | 1.0 | amide p aminobenzenesulfonamide phenylmethylsulfonyl fluoride n 1 naphthyl ethylenediamine dihydrochloride bathocuproinedisulfonic acid monoclonal anti bcl 2 clone bcl 2 100 monoclonal anti p53 clone bp53 12 and cytochrome c horse heart were obtained from sigma. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | obenzenesulfonamide phenylmethylsulfonyl fluoride n 1 naphthyl ethylenediamine dihydrochloride bathocuproinedisulfonic acid monoclonal anti bcl 2 clone bcl 2 100 monoclonal anti p53 clone bp53 12 and cytochrome c horse heart were obtained from sigma. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | ac devd amc caspase 3 inhibitor i ac devd cho hoechst 33342 diethylamine nonoate nono and 3 [ _amp_#177; e ethyl 2' [ e hydroxyimino] 5 nitro 3 exenecarbamoyl]pyridine nor 4 were purchased from calbiochem. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | anti cytochrome c monoclonal antibody clone 7h8.2c12 was from pharmingen san diego ca . |
| 11180 | SOD2 | superoxide dismutase 2, mitochondrial | mn sod | 1.0 | transfected cells had the same antioxidant pattern such as of mn sod as sh sy5y cells. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | cytosolic cytochrome c determination cells detached and attached were washed with pbs and collected by centrifugation at 700 _amp_#215; g for 5 min at 4 degreec. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | purified cytochrome c from horse heart was used as standard. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | purified mouse anti cytochrome c monoclonal antibody was used as primary antibody 1:5000 . |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | p53 p21 and bcl 2 proteins detection cell pellet was resuspended in lysis buffer containing 10 m m tris/hcl ph 7.4 5 m m edta 150 m m nacl 0.5% octyphenoxy polyethoxyethanol igepal ca 630 sigma and protease inhibitors |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | sates were then centrifuged at 14 000 _amp_#215; g for 15 min at 4 degreec and supernatants were removed and stored at 80 degreec. 50 microg of proteins were loaded on a 10 for p53 or 12% for p21 and bcl 2 polyacrylamide gel. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | monoclonal anti bcl 2 1:5000 monoclonal anti p53 1:5000 or polyclonal anti p21 1:500 was used as primary antibodies. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | caspase 3 activation cells attached and detached were washed and collected by centrifugation at 700 _amp_#215; g for 5 min at 4 degreec. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | supernatants were used for caspase 3 assay in lysis buffer containing the specific substrate ac devd amc 12 micro m at 30 degreec. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | specific activity was demonstrated by the use of the caspase 3 inhibitor i ac devd cho that completely impedes the development of fluorescence due to the cleaved product. |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | cu zn superoxide dismutase activity cells attached and detached were washed and collected by centrifugation. |
| 11180 | SOD2 | superoxide dismutase 2, mitochondrial | mn sod | 1.0 | under these conditions the activity of mn sod is almost fully inhibited 33 . |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | cytochrome c oxidase activity cells attached and detached were washed and collected by centrifugation. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | lysates were used for cytochrome c oxidase activity. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | the activity was measured spectrophotometrically monitoring the oxidation of cytochrome c horse heart which had previously been reduced by treatment with excess ascorbate followed by passage on sephadex g 25 resin amersham pharmacia biotech . |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | activity was expressed as micromoles of cytochrome c oxidized per min mg protein using an extinction coefficient of 27.6 m m cm . |
| 2295 | CP | ceruloplasmin (ferroxidase) | ceruloplasmin | 1.0 | the reaction between gsh and no has been suggested to be catalyzed by ceruloplasmin a multicopper oxidase involved in iron metabolism. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | it has been demonstrated that p21 cleavage could be reproduced in extracts prepared from irradiated cells or by recombinant caspase 3 suggesting that a caspase like activity is responsible for this cleavage. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | nuclear apoptosis is preceded by the disruption of the mitochondrial transmembrane potential which in turn can facilitate the release of pro apoptotic proteins such as cytochrome c through the opening of the mitochondrial permeability transition pore 54 . |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | in a previous report regarding another cell model system undergoing apoptosis 56 we demonstrated that cytochrome c release from mitochondria followed oxidative stress consequent to glutathione depletion. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | furthermore this phenomenon could be observed in healthy cells as well suggesting that cytochrome c release is part of a more general mechanism related to redox unbalance. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | in contrast to this hypothesis in the present report gsno treatment led to cytochrome c release with the maintenance of high intracellular glutathione concentration. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | moreover in the g93a cells a small level of released cytochrome c was also detected in the untreated cells indicating that these cells in culture may spontaneously die by apoptosis. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | taken together these results point to a mechanism for cytochrome c release different from that related to glutathione depletion. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | a specific marker of no toxicity was the decrease of cytochrome c oxidase activity which paralleled the extent of apoptosis. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | an alternative model for protein export from mitochondria depends on bax and/or bid two members of the bcl 2 protein family 57 . |
| 992 | BCL2L1 | BCL2-like 1 | bcl xl | 1.0 | it involves the translocation of bid or bax from cytosol to mitochondria where they may possibly form a pore as shown for the related protein bcl xl 58 . |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | in our experiments bcl 2 was down regulated by the gsno treatment in sh sy5y cells whereas the resistance of wt cells to apoptosis was associated with a higher level of this protein in the treated cells. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | since they were the most prone to no induced apoptosis we propose a causative role for bcl 2 in the increased susceptibility of g93a cells to apoptosis. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | it has been suggested that the antiapoptotic effects of bcl 2 be at least in part due to its antioxidant activity 59 . |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | t known it is established that the cytosolic steady state of reactive oxygen species which are potentially threatening species and modulators of physiological signal transduction 60 is increased when bcl 2 is down regulated. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | a vicious circle including down regulation of bcl 2 and deviating redox activity of copper in the g93a mutant may be the amplification factor leading to ros unbalance in g93a cells and may explain how and why an als mutant cause a pro apoptotic respon |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | a cytochrome c released in the cytosol was detected by western blotting using a monoclonal antibody as described under "experimental procedures." 50 microg of cytosolic protein were loaded on each lane. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | lane 1 sh sy5y cells; lane 2 gsno treated; lane 3 g93a cells; lane 4 gsno treated; lane 5 wt cells; lane 6 gsno treated; lane 7 purified cytochrome c. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | b caspase 3 activity was measured fluorometrically with ac devd amc as substrate. |
| 5173 | HRAS | v-Ha-ras Harvey rat sarcoma viral oncogene homolog | p21 protein | 1.0 | immunoreactive p53 protein c and immunoreactive p21 protein d were measured by western blotting using monoclonal antibody as detailed under "experimental procedures." 50 microg protein of cell extracts was applied to each lane. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | bcl 2 content of sh sy5y cells and effects on it by cu zn sod and gsno treatment. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | bcl 2 immunoreactive protein was detected by western blotting using a monoclonal antibody as described under "experimental procedures." 50 microg protein of cell extracts was loaded on each lane. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | b activity of cytochrome c oxidase. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | spectrophotometric determination of cytochrome c oxidase activity was performed as described under "experimental procedures." |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl2 | 1.0 | apoptotic markers: cytochrome c release caspase activation p53 accumulation p21 increase and bcl2 down regulation to examine the sequence of events occurring upon gsno toxicity we measured some of the molecular markers of apoptotic cell death at 48 h from the apoptogenic stimulus. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | apoptotic markers: cytochrome c release caspase activation p53 accumulation p21 increase and bcl2 down regulation to examine the sequence of events occurring upon gsno toxicity we measured some of the molecular markers of apoptotic |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | cytosolic extracts were prepared under conditions that keep mitochondria intact and cytosolic cytochrome c protein levels were measured by immunoblot analysis. |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | the cytosol from untreated sh sy5y and wt cells contained no detectable amounts of cytochrome c whereas untreated g93a cells had a small level of released cytochrome c probably as result of spontaneous apoptotic death fig 3 a . |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | cytosolic cytochrome c accumulated after gsno treatment; the release was highest for g93a cells and almost at the limit of detection for wt cells. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | proteolytic activity associated to caspases was measured by testing the cytosolic extracts for their ability to cleave the fluorimetric substrate ac devd amc which is specific for caspase 3. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | caspase 3 activation is important in the cascade of proteolytic events of apoptosis in that it is considered one of the points of no return in the process leading to cell destruction 40 . |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | caspase 3 activation followed the same trend as cytochrome c release with higher activity in the cytosolic extracts of g93a cells than in sh sy5y and wt cells fig 3 b . |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | caspase 3 activation followed the same trend as cytochrome c release with higher activity in the cytosolic extracts of g93a cells than in sh sy5y and wt cells fig 3 b . |
| 13716 | C4orf6 | chromosome 4 open reading frame 6 | expressed in neuroblastoma | 1.0 | fig. 3 c shows that the p53 protein was stably expressed in neuroblastoma cells to a varying extent; in particular wt cells showed a lower p53 protein level with respect to sh sy5y cells and g93a cells which showed the higher level. |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | bcl 2 protein modulation is responsible for cell survival or suicide; constitutive expression of high bcl 2 protein levels by transfection experiments has proven that bcl 2 or related family members can protect cells from no mediated apoptosis 44 45 . |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | in our experiments as evidenced by western blot analysis untreated sh sy5y and wt cells express high levels of bcl 2 protein whereas g93a cells show a much lower content fig 4 . 48 h of gsno treatment decreased bcl 2 expression although to a different extent; in particular bcl 2 content of g93a cells was at the lim |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | protein whereas g93a cells show a much lower content fig 4 . 48 h of gsno treatment decreased bcl 2 expression although to a different extent; in particular bcl 2 content of g93a cells was at the limit of detection. |
| 31395 | COX8B | cytochrome c oxidase, subunit 8B pseudogene | cytochrome c oxidase | 1.0 | among the above mentioned molecular target of no toxicity the hemoprotein cytochrome c oxidase appears to be the only one strongly affected by gsno insult and protected by native cu zn sod fig 7 b . |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | nono diethylamine nonoate; nor 4 3 [ _amp_#177; e ethyl 2' [ e hydroxyimino] 5 nitro 3 exenecarbamo yl]pyridine; dcf da 2' 7' dichlorodihydrofluorescein diacetate; ac devd amc ac asp glu val asp amc caspase 3 substrate ii fluorogenic; ac devd cho ac asp glu val asp cho caspase 3 inhibitor i; ros reactive oxygen species; fals familial amyotrophic lateral sclerosis; pipes piperazine n n ' bis[2 ethanesulfon |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | substrate ii fluorogenic; ac devd cho ac asp glu val asp cho caspase 3 inhibitor i; ros reactive oxygen species; fals familial amyotrophic lateral sclerosis; pipes piperazine n n ' bis[2 ethanesulfonic acid]; chaps 3 [cholamindopropyl dimethylammonio] 1 propanesulfonate |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | the abbreviations used are: no nitric oxide; wt sh sy5y transfected with wild type cu zn sod; g93a sh sy5y transfected with mutant g93a cu zn sod; cu zn sod copper zinc superoxide dismutase; gsno s nitrosoglutathione; nono diethylamine nonoate; nor 4 3 [ _amp_#177; e ethyl 2' [ e hydroxyimino] 5 nitro 3 exenecarbamo yl]pyridine; dcf da 2' 7' dichlorodihydrofluorescein diacetate; ac devd |
| 990 | BCL2 | B-cell CLL/lymphoma 2 | bcl 2 | 1.0 | cytochrome c group|nitroso compounds|proto oncogene proteins c bcl 2|tumor suppressor protein p53|nitric oxide|s nitrosoglutathione|glutathione|superoxide dismutase|caspases|oncogene protein p2| |
| 19986 | CYCS | cytochrome c, somatic | cytochrome c | 1.0 | cytochrome c group|nitroso compounds|proto oncogene proteins c bcl 2|tumor suppressor protein p53|nitric oxide|s nitrosoglutathione|glutathione|superoxide dismutase|caspases|oncogene protein p2| |