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4.
Biochim Biophys Acta Mol Cell Res ; 1865(8): 1046-1059, 2018 08.
Article in English | MEDLINE | ID: mdl-29694914

ABSTRACT

GSK3ß interacting protein (GSKIP) is a naturally occurring negative regulator of GSK3ß and retains both the Protein Kinase A Regulatory subunit binding (PKA-RII) domain and GSK3ß interacting domain. Of these two domains, we found that PKA-RII is required for forming a working complex comprising PKA/GSKIP/GSK3ß/Drp1 to influence phosphorylation of Drp1 Ser637. In this study, bioinformatics and experimental explorations re-analyzing GSKIP's biofunctions suggest that the evolutionarily conserved Domain of Unknown Function (DUF727) is an ancestral prototype of GSKIP in prokaryotes, and acquired the C-terminal GSK3ß binding site (tail) in invertebrates except for Saccharomyces spp., after which the N-terminal PKA-RII binding region (head) evolved in vertebrates. These two regions mutually influence each other and modulate GSKIP binding to GSK3ß in yeast two-hybrid assays and co-immunoprecipitation. Molecular modeling showed that mammalian GSKIP could form a dimer through the L130 residue (GSK3ß binding site) rather than V41/L45 residues. In contrast, V41/L45P mutant facilitated a gain-of-function effect on GSKIP dimerization, further influencing binding behavior to GSK3ß compared to GSKIP wild-type (wt). The V41/L45 residues are not only responsible for PKA RII binding that controls GSK3ß activity, but also affect dimerization of GSKIP monomer, with net results of gain-of-function in GSKIP-GSK3ß interaction. In addition to its reported role in modulating Drp1, Ser637 phosphorylation caused mitochondrial elongation; we postulated that GSKIP might be involved in the Wnt signaling pathway as a scavenger to recruit GSK3ß away from the ß-catenin destruction complex and as a competitor to compete for GSK3ß binding, resulting in accumulation of S675 phosphorylated ß-catenin.


Subject(s)
Repressor Proteins/chemistry , Repressor Proteins/metabolism , Wnt Signaling Pathway , Binding Sites , Computational Biology , Cyclic AMP-Dependent Protein Kinases/metabolism , Dynamins , Evolution, Molecular , GTP Phosphohydrolases/chemistry , GTP Phosphohydrolases/metabolism , Glycogen Synthase Kinase 3 beta/metabolism , HEK293 Cells , Humans , Microtubule-Associated Proteins/chemistry , Microtubule-Associated Proteins/metabolism , Mitochondrial Proteins/chemistry , Mitochondrial Proteins/metabolism , Models, Molecular , Phosphorylation , Phylogeny , Protein Binding , Protein Domains , Protein Multimerization , Repressor Proteins/genetics , Serine/chemistry , Two-Hybrid System Techniques
5.
Cell Cycle ; 14(17): 2764-76, 2015.
Article in English | MEDLINE | ID: mdl-26114227

ABSTRACT

We previously reported that Aurora-A and the hNinein binding protein AIBp facilitate centrosomal structure maintenance and contribute to spindle formation. Here, we report that AIBp also interacts with Plk1, raising the possibility of functional similarity to Bora, which subsequently promotes Aurora-A-mediated Plk1 activation at Thr210 as well as Aurora-A activation at Thr288. In kinase assays, AIBp acts not only as a substrate but also as a positive regulator of both Aurora-A and Plk1. However, AIBp functions as a negative regulator to block phosphorylation of hNinein mediated by Aurora-A and Plk1. These findings suggest a novel AIBp-dependent regulatory machinery that controls mitotic entry. Additionally, knockdown of hNinein caused failure of AIBp to target the centrosome, whereas depletion of AIBp did not affect the localization of hNinein and microtubule nucleation. Notably, knockdown of AIBp in HeLa cells impaired both Aurora-A and Plk1 kinase, resulting in phenotypes with multiple spindle pole formation and chromosome misalignment. Our data show that depletion of AIBp results in the mis-localization of TACC3 and ch-TOG, but not CEP192 and CEP215, suggesting that loss of AIBp dominantly affects the Aurora-A substrate to cause mitotic aberrations. Collectively, our data demonstrate that AIBp contributes to mitotic entry and bipolar spindle assembly and may partially control localization, phosphorylation, and activation of both Aurora-A and Plk1 via hNinein during mitotic progression.


Subject(s)
Aurora Kinase A/metabolism , Carrier Proteins/physiology , Cell Cycle Proteins/metabolism , Mitosis/physiology , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins/metabolism , Spindle Apparatus/metabolism , Aurora Kinase A/genetics , Cell Cycle Proteins/genetics , DNA-Binding Proteins , HEK293 Cells , HeLa Cells , Humans , Protein Serine-Threonine Kinases/genetics , Proto-Oncogene Proteins/genetics , Spindle Apparatus/genetics , Polo-Like Kinase 1
6.
Biochim Biophys Acta ; 1853(8): 1796-807, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25920809

ABSTRACT

GSK3ß binding of GSKIP affects neurite outgrowth, but the physiological significance of PKA binding to GSKIP remains to be determined. We hypothesized that GSKIP and GSK3ß mediate cAMP/PKA/Drp1 axis signaling and modulate mitochondrial morphology by forming a working complex comprising PKA/GSKIP/GSK3ß/Drp1. We demonstrated that GSKIP wild-type overexpression increased phosphorylation of Drp1 S637 by 7-8-fold compared to PKA kinase-inactive mutants (V41/L45) and a GSK3ß binding-defective mutant (L130) under H2O2 and forskolin challenge in HEK293 cells, indicating that not only V41/L45, but also L130 may be involved in Drp1-associated protection of GSKIP. Interestingly, silencing either GSKIP or GSK3ß but not GSK3α resulted in a dramatic decrease in Drp1 S637 phosphorylation, revealing that both GSKIP and GSK3ß are required in this novel PKA/GSKIP/GSK3ß/Drp1 complex. Moreover, overexpressed kinase-dead GSK3ß-K85R, which retains the capacity to bind GSKIP, but not K85M which shows total loss of GSKIP-binding, has a higher Drp1 S637 phosphorylation similar to the GSKIP wt overexpression group, indicating that GSK3ß recruits Drp1 by anchoring rather than in a kinase role. With further overexpression of either V41/L45P or the L130P GSKIP mutant, the elongated mitochondrial phenotype was lost; however, ectopically expressed Drp1 S637D, a phosphomimetic mutant, but not S637A, a non-phosphorylated mutant, restored the elongated mitochondrial morphology, indicating that Drp1 is a downstream effector of direct PKA signaling and possibly has an indirect GSKIP function involved in the cAMP/PKA/Drp1 signaling axis. Collectively, our data revealed that both GSKIP and GSK3ß function as anchoring proteins in the cAMP/PKA/Drp1 signaling axis modulating Drp1 phosphorylation.


Subject(s)
Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclic AMP/metabolism , GTP Phosphohydrolases/metabolism , Glycogen Synthase Kinase 3/physiology , Microtubule-Associated Proteins/metabolism , Mitochondria/metabolism , Mitochondrial Proteins/metabolism , Repressor Proteins/physiology , Cells, Cultured , Dynamins , GTP Phosphohydrolases/genetics , Glycogen Synthase Kinase 3/metabolism , HEK293 Cells , HeLa Cells , Humans , Microtubule-Associated Proteins/genetics , Mitochondria/genetics , Mitochondrial Dynamics/physiology , Mitochondrial Proteins/genetics , Phosphorylation , Repressor Proteins/metabolism , Signal Transduction/genetics
7.
Int J Oncol ; 46(3): 1304-16, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25586056

ABSTRACT

Bcl2L12 as a new member of the Bcl2 family, which contains a BH2 domain and shares a lower amino acid similarity with other Bcl2 family proteins. Bcl2L12 is reported to be involved in apoptosis regulation, but this role remains controversial in different cancer type. Temozolomide (TMZ) is currently used to intervene glioma multiforme (GBM), but an acquired chemotherapeutic resistance maybe occurred due to undesired autophagy. Previous studies uncovered that Bcl2L12 may interact with Bcl-xL and may harbor a BH3-like domain. Therefore, we investigated whether this BH3-like domain is responsible for the Bcl2L12 anti-apoptotic property. Moreover, we tested whether ABT-737, a BH3 mimetic agent, can be combined with TMZ to treat GBM. We aligned Bcl2L12 with Bcl2 family members, compared interacting pattern of BH3 domain and their protein 3D structure. We identified that Bcl2L12 interacts with Bcl-xL and Bcl2 in yeast two-hybrid system. Bcl2L12192-220 was a minimal region for Bcl2L12-Bcl-xL interaction. Five-point mutations with respect to hydrophobic and charge residues were generated to test whether they are the key residue of BH3-like domain. Our data showed that both h1 (L213) and h2 residue (L217) are essential for Bcl2L12 interacting with Bcl2 family proteins. Ectopically expressed h1 or h2 mutant in U87MG cell line resulted in reactivation of cleaved-PARP, caspase-3 and cytochrome c releasing compared to Bcl2L12 wt group. Implementing ABT-737 combined with TMZ provided a superior effect on apoptosis induction in Bcl2L12 wt group, which effectively reactivated apoptotic markers. Altogether, our findings indicated that Bcl2L12 retains a BH3-like domain, which is important for the Bcl2L12 anti-apoptotic property and TMZ-induced autophagy. Our results basically support the idea of using ABT-737 to counteract the anti-apoptotic role of Bcl2L12 and sensitize drug response of the GBM cells to TMZ.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Apoptosis/genetics , Autophagy/drug effects , Biphenyl Compounds/administration & dosage , Dacarbazine/analogs & derivatives , Glioma/drug therapy , Muscle Proteins/physiology , Nitrophenols/administration & dosage , Proto-Oncogene Proteins c-bcl-2/physiology , Sulfonamides/administration & dosage , Dacarbazine/administration & dosage , Dacarbazine/pharmacology , Drug Resistance, Neoplasm/drug effects , Drug Resistance, Neoplasm/genetics , Glioma/genetics , Glioma/pathology , Humans , Models, Molecular , Muscle Proteins/antagonists & inhibitors , Muscle Proteins/chemistry , Peptide Fragments/chemistry , Piperazines/administration & dosage , Protein Binding , Protein Structure, Tertiary , Proto-Oncogene Proteins/chemistry , Proto-Oncogene Proteins c-bcl-2/antagonists & inhibitors , Proto-Oncogene Proteins c-bcl-2/chemistry , Proto-Oncogene Proteins c-bcl-2/metabolism , Temozolomide , Tumor Cells, Cultured , bcl-X Protein/metabolism
8.
Neurol Res ; 36(8): 746-51, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24620973

ABSTRACT

OBJECTIVES: The purpose was to evaluate the association of Aurora A and gamma-tubulin expression with disease characteristics and survival in patients with astrocytoma. METHODS: This is a retrospective study of patients who had surgical specimens that were pathologically diagnosed as astrocytoma. The expression level of Aurora A and gamma-tubulin in tumor tissue was evaluated by immunohistochemistry. Clinical information, Karnofsky performance status scale, and survival status of patients were collected. RESULTS: We found that high protein levels of gamma-tubulin or Aurora A were associated with patients 45 years of age, high tumor grade, more advanced non-fully resectable tumors, and poorer survival status. The survival time for patients whose tumors had high gamma-tubulin and Aurora A expression was about 12 months compared with approximately 41 months for patients with low levels of expression of these proteins. Poor patient performance status following resection was also associated with high levels of gamma-tubulin and Aurora A expression. DISCUSSION: The expression levels of gamma-tubulin or Aurora A kinase were associated with patients' age, astrocytoma grade, respectability, as well as patient survival and performance. These findings support the idea that these factors may potentially be important prognostic indicators for patients with astrocytomas.


Subject(s)
Astrocytoma/metabolism , Astrocytoma/mortality , Aurora Kinase A/metabolism , Brain Neoplasms/metabolism , Brain Neoplasms/mortality , Tubulin/metabolism , Adult , Astrocytoma/enzymology , Brain Neoplasms/enzymology , Female , Humans , Karnofsky Performance Status , Male , Middle Aged , Retrospective Studies , Survival Analysis
9.
Childs Nerv Syst ; 29(11): 2051-5, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23632690

ABSTRACT

PURPOSE: The prognosis of children with low-grade cerebellar astrocytoma who have partial resection of tumor is largely unpredictable. The purpose of this study was to review the long-term outcome of such patients. METHODS: The medical charts, imaging findings, operative notes, histopathological reports, and survival times of 12 patients with cerebellar astrocytoma were reviewed. RESULTS: Five patients had total resection and seven had partial resection. Nine patients had grade I histology and three patients had grade II. Follow-up duration ranged from 3 to 25 years. Among the seven patients with residual tumor, five had tumor progression, one had arrested tumor growth, and one had spontaneous tumor regression. Five patients with partial resection received radiotherapy and three had malignant transformation of tumor during follow-up. Six patients, including five who had partial resection, underwent a second operation. One patient with partial resection died of pneumonia 23 years after surgery. CONCLUSIONS: Patients with complete tumor resection had a better prognosis than patients with partial resection. For patients with partial resection, we recommend a "wait and see" policy with surveillance using MRI. The phenomenon of arrested tumor growth and spontaneous tumor regression in patients with cerebellar astrocytoma who have subtotal resection warrants further study.


Subject(s)
Astrocytoma , Cerebellar Neoplasms , Neoplasm Recurrence, Local , Neoplasm, Residual , Neurosurgical Procedures/methods , Adolescent , Astrocytoma/pathology , Astrocytoma/surgery , Cerebellar Neoplasms/pathology , Cerebellar Neoplasms/surgery , Child , Child, Preschool , Disease Progression , Female , Follow-Up Studies , Humans , Male , Neoplasm Grading , Neoplasm Recurrence, Local/pathology , Neoplasm Recurrence, Local/surgery , Neoplasm, Residual/pathology , Neoplasm, Residual/surgery , Neurosurgical Procedures/standards , Prognosis , Reoperation , Retrospective Studies , Treatment Outcome
10.
Psychiatry Res ; 209(3): 424-30, 2013 Oct 30.
Article in English | MEDLINE | ID: mdl-23598059

ABSTRACT

This study assessed facial emotion recognition abilities in subjects with paranoid and non-paranoid schizophrenia (NPS) using signal detection theory. We explore the differential deficits in facial emotion recognition in 44 paranoid patients with schizophrenia (PS) and 30 non-paranoid patients with schizophrenia (NPS), compared to 80 healthy controls. We used morphed faces with different intensities of emotion and computed the sensitivity index (d') of each emotion. The results showed that performance differed between the schizophrenia and healthy controls groups in the recognition of both negative and positive affects. The PS group performed worse than the healthy controls group but better than the NPS group in overall performance. Performance differed between the NPS and healthy controls groups in the recognition of all basic emotions and neutral faces; between the PS and healthy controls groups in the recognition of angry faces; and between the PS and NPS groups in the recognition of happiness, anger, sadness, disgust, and neutral affects. The facial emotion recognition impairment in schizophrenia may reflect a generalized deficit rather than a negative-emotion specific deficit. The PS group performed worse than the control group, but better than the NPS group in facial expression recognition, with differential deficits between PS and NPS patients.


Subject(s)
Affective Symptoms/etiology , Emotions , Recognition, Psychology/physiology , Schizophrenia, Paranoid/complications , Schizophrenia/complications , Schizophrenic Psychology , Adult , Facial Expression , Female , Humans , Male , Neuropsychological Tests , Psychiatric Status Rating Scales , Signal Detection, Psychological , Young Adult
11.
Cytokine ; 61(2): 413-8, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23260996

ABSTRACT

Transforming growth factor-beta 1 (TGF-ß1) has been reported to be a possible marker for a number of tumors, including brain tumors. The aim of this study was to measure the plasma levels of TGF-ß1 in patients with low- and high-grade astrocytomas before and after surgery. This prospective study included 14 patients with low-grade astrocytomas and 25 with high-grade astrocytomas who underwent tumor removal and 13 controls (patients who underwent cranioplasty for skull bone defects). Plasma levels of TGF-ß1 were measured in all subjects using enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve analysis showed that when the level of TGF-ß1 before tumor removal was ≥ 2.52 ng/ml, astrocytoma was predicted with a sensitivity of 94.9% and specificity of 100%. The mean plasma level of TGF-ß1 in both the low-grade and high-grade astrocytoma groups significantly decreased after tumor removal (p<0.05); there was no significant change in TGF-ß1 plasma level of the controls following surgery. Patients with high-grade astrocytomas had a significantly higher mortality rate than patients with low-grade astrocytomas (p=0.019) and significantly shorter survival (p=0.008). A positive correlation between TGF-ß1 level after tumor removal and tumor volume was only found in the high-grade astrocytoma group (γ=0.597, p=0.002). The findings show that plasma TGF-ß1 level was increased in patients with low-grade and high-grade astrocytoma, and that the levels significantly decreased after tumor removal in both groups. The results provide additional evidence that TGF-ß1 might be useful as a tumor marker for astrocytomas.


Subject(s)
Astrocytoma/blood , Astrocytoma/surgery , Transforming Growth Factor beta1/blood , Adolescent , Adult , Aged , Astrocytoma/pathology , Case-Control Studies , Child , Child, Preschool , Female , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Neoplasm Grading , ROC Curve , Survival Analysis , Treatment Outcome , Young Adult
12.
PLoS One ; 7(11): e49112, 2012.
Article in English | MEDLINE | ID: mdl-23185298

ABSTRACT

Multiple phosphorylation sites of Drp1 have been characterized for their functional importance. However, the functional consequence of GSK3beta-mediated phosphorylation of Drp1 remains unclear. In this report, we pinpointed 11 Serine/Threonine sites spanning from residue 634~736 of the GED domain and robustly confirmed Drp1 Ser693 as a novel GSK3beta phosphorylation site. Our results suggest that GSK3beta-mediated phosphorylation at Ser693 does cause a dramatic decrease of GTPase activity; in contrast, GSK3beta-mediated phosphorylation at Ser693 appears not to affect Drp1 inter-/intra-molecular interactions. After identifying Ser693 as a GSK3beta phosphorylation site, we also determined that K679 is crucial for GSK3beta-binding, which strongly suggests that Drp1 is a novel substrate for GSK3beta. Thereafter, we found that overexpressed S693D, but not S693A mutant, caused an elongated mitochondrial morphology which is similar to that of K38A, S637D and K679A mutants. Interestedly, using H89 and LiCl to inhibit PKA and GSK3beta signaling, respectively, it appears that a portion of the elongated mitochondria switched to a fragmented phenotype. In investigating the biofunctionality of phosphorylation sites within the GED domain, cells overexpressing Drp1 S693D and S637D, but not S693A, showed an acquired resistance to H(2)O(2)-induced mitochondrial fragmentation and ensuing apoptosis, which affected cytochrome c, capase-3, -7, and PARP, but not LC3B, Atg-5, Beclin-1 and Bcl2 expressions. These results also showed that the S693D group is more effective in protecting both non-neuronal and neuronal cells from apoptotic death than the S637D group. Altogether, our data suggest that GSK3beta-mediated phosphorylation at Ser693 of Drp1 may be associated with mitochondrial elongation via down-regulating apoptosis, but not autophagy upon H(2)O(2) insult.


Subject(s)
Glycogen Synthase Kinase 3/metabolism , Mitochondria/enzymology , Mitochondria/pathology , Oxidative Stress , Amino Acid Sequence , Apoptosis/drug effects , Autophagy/drug effects , Dynamins/chemistry , Dynamins/metabolism , GTP Phosphohydrolases/metabolism , Glycogen Synthase Kinase 3 beta , HEK293 Cells , HeLa Cells , Humans , Hydrogen Peroxide/pharmacology , Hydrolysis/drug effects , Lysine/metabolism , Mitochondria/drug effects , Mitochondrial Dynamics/drug effects , Models, Biological , Molecular Sequence Data , Mutant Proteins/chemistry , Mutant Proteins/metabolism , Oxidative Stress/drug effects , Phosphorylation/drug effects , Phosphoserine/metabolism , Protein Binding/drug effects , Protein Structure, Tertiary , Protein Transport/drug effects , Two-Hybrid System Techniques
13.
Biofactors ; 38(6): 470-7, 2012.
Article in English | MEDLINE | ID: mdl-23047184

ABSTRACT

Dysregulated centrosomal expression has been observed in high grade gliomas. Thus, this study aimed to examine the expression of Aurora family kinase and various centrosomal proteins, including centrin, γ-tubulin, and hNinein isoforms, in human brain tumors, including 29 meningiomas, 34 astrocytomas, 6 pituitary adenomas, and 6 metastatic tumors. mRNA expression was evaluated using reverse transcription polymerase chain reaction. The role of hNinein isoform 6 expression in cell differentiation was assessed in BrdU-treated IMR-32 cells. Differential expression of centrosomal proteins of brain tumors and cell lines was observed. Specifically, centrin 2 and centrin 3 expression levels were classified as moderate or abundant in >97% of samples in the meningioma group, 63% of astrocytomas, >83% of metastatic and pituitary tumors. Alternatively, hNinein isoform 6 expression was only detected in normal brain and astrocytoma tumors (17/34); however, it was not expressed in meningioma (0/29), metastatic tumors (0/6) (P < 0.001). Of the six neuroblastoma cell lines analyzed only IMR-32 cells expressed hNinein isoform 6. Furthermore, downregulated expression of hNinein isoform 6 and upregulation of γ-tubulin was correlated to astrocytoma tumor grade (P < 0.001). Increased hNinein isoform 6 mRNA expression was observed in response to BrdU treatment, and its expression was greater in teratomas as compared to embryonic stem cells. Further studies are necessary to determine if hNinein isoform 6 functions as a tumor-suppressor gene in brain tumors. Differential centrosomal protein expression may result in altered centrosome function that is observed the in progression of various brain tumors.


Subject(s)
Brain Neoplasms/metabolism , Cell Differentiation , Centrosome/metabolism , Cytoskeletal Proteins/genetics , Cytoskeletal Proteins/metabolism , Microtubule-Associated Proteins/genetics , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Brain/metabolism , Brain/pathology , Brain Neoplasms/genetics , Humans , Microtubule-Associated Proteins/metabolism , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA, Messenger/metabolism , Tubulin/genetics , Tubulin/metabolism
14.
Kaohsiung J Med Sci ; 28(6): 316-21, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22632886

ABSTRACT

Transforming growth factor (TGF) ß1 may be a candidate for a serologic tumor marker. In this study, the plasma levels of TGF-ß1 in patients with brain tumors were measured using enzyme-linked immunosorbent assay before and after tumor removal. Patients were divided into four groups, the control group and the benign, malignant, and metastatic brain tumor groups. All brain tumor groups showed significant increases in the levels of TGF-ß1 before tumor removal (6.36 ± 3.94, 17.0 ± 9.7, and 12.2 ± 10.3 ng/ml for the benign, malignant, and metastatic groups, respectively). When compared with the results obtained in the control group (1.12 ± 0.74 ng/ml), significant decreases in TGF-ß1 concentrations after total tumor removal were found in both the benign and malignant brain tumor groups (2.55 ± 2.00 and 8.93 ± 5.73 ng/ml, respectively; p = 0.0001 and p = 0.003, respectively). On the other hand, plasma TGF-ß1 levels in the metastatic brain tumor group showed a slight but significant increase (14.7 ± 9.3 ng/ml, p = 0.035) after tumor removal. In a case of low-grade astrocytoma, plasma levels of TGF-ß1 were found to be 3.6 and 1.1 ng/ml before and after tumor removal, respectively. However, recurrent tumor was noted in this patient 7 months later, and the levels of TGF-ß1 were 26.2 and 8.4 ng/ml before and after the second operation, respectively. The data show that plasma TGF-ß1 was elevated in the circulation of patients with brain tumors and that significant decreases in TGF-ß1 levels were observed after the removal of benign and malignant tumors. The results also suggest that TGF-ß1 may be a useful serologic marker for brain tumors.


Subject(s)
Biomarkers, Tumor/blood , Brain Neoplasms/blood , Brain Neoplasms/surgery , Transforming Growth Factor beta1/blood , Adolescent , Adult , Aged , Brain Neoplasms/pathology , Child , Female , Humans , Male , Middle Aged , Young Adult
15.
Shock ; 38(2): 203-12, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22576001

ABSTRACT

Alterations in the activity of vascular K channels are commonly associated with abnormalities in cerebral vascular function after subarachnoid hemorrhage (SAH). Subarachnoid hemorrhage-induced vasospasm remains incompletely understood; nevertheless, activation of K channels may be of benefit in relieving spastic constriction. This study was to examine whether the vasodilators KMUP-1 and pinacidil, a KATP-channel opener, have the ability to prevent SAH-induced vasospasm via the large-conductance Ca-activated K (BKCa) channels in cerebral arteries. Rats were divided into four groups including sham-operated, SAH, KMUP-1 treated, and pinacidil treated. Subarachnoid hemorrhage rats were induced by injecting autologous blood into cisterna magna, and then KMUP-1 or pinacidil (1 mg/kg) was injected intraperitoneally 1 and 24 h after SAH. Behavioral tests were assessed on day 2 after SAH before the rats were killed. Cerebral myocytes were enzymatically isolated from rat basilar arteries and used to monitor BKCa-channel activities. In isolated basilar arteries, KMUP-1-treated and pinacidil-treated rats showed normalized vascular reactivity. In whole-cell recordings, BKCa currents were attenuated in SAH rats compared with sham-operated rats. In inside-out patches, the conductance and voltage sensitivity of single BKCa-channels were unchanged among the four groups. In contrast, SAH rats showed markedly decreased BKCa-channel activity and ß1-subunit expression associated Ca sensitivity that was further confirmed by immunofluorescence staining and Western blotting. Subarachnoid hemorrhage-induced deficits in motor function and BKCa-channel inhibition were improved by KMUP-1-treated and pinacidil-treated rats. In addition, SAH appears to modify BKCa-channel calcium sensitivity. KMUP-1 and pinacidil prevent SAH-induced vasospasm at least in part by the restoration of BKCa-channel activities.


Subject(s)
Large-Conductance Calcium-Activated Potassium Channels/drug effects , Pinacidil/pharmacology , Piperidines/pharmacology , Subarachnoid Hemorrhage/complications , Vasodilator Agents/pharmacology , Vasospasm, Intracranial/prevention & control , Xanthines/pharmacology , Animals , Basilar Artery/drug effects , Down-Regulation , Male , Muscle, Smooth, Vascular , Psychomotor Performance/physiology , Rats , Rats, Sprague-Dawley , Vasoconstriction/drug effects , Vasospasm, Intracranial/etiology
16.
Psychiatry Res ; 200(2-3): 928-32, 2012 Dec 30.
Article in English | MEDLINE | ID: mdl-22503384

ABSTRACT

The Chinese Facial Emotion Recognition Database (CFERD), a computer-generated three-dimensional (3D) paradigm, was developed to measure the recognition of facial emotional expressions at different intensities. The stimuli consisted of 3D colour photographic images of six basic facial emotional expressions (happiness, sadness, disgust, fear, anger and surprise) and neutral faces of the Chinese. The purpose of the present study is to describe the development and validation of CFERD with nonclinical healthy participants (N=100; 50 men; age ranging between 18 and 50 years), and to generate normative data set. The results showed that the sensitivity index d' [d'=Z(hit rate)-Z(false alarm rate), where function Z(p), p∈[0,1]], for all emotions was 0.94. The emotion was more readily detected in happiness, and less easily detected in surprise and sadness. In general, this study replicated the previous findings on the recognition accuracy of emotional expression with the Westerner faces. However, our paradigm extends the previous work by including a wider sensitivity range to differentiate subtle perception of emotion intensities. The CFERD will be a useful tool for emotion recognition assessment in affective neurosciences research, especially for the Chinese and cross-cultural studies.


Subject(s)
Emotions/physiology , Facial Expression , Recognition, Psychology/physiology , Social Perception , Adolescent , Adult , China , Databases, Factual , Female , Humans , Male , Middle Aged , Photic Stimulation/methods
17.
Cell Cycle ; 11(3): 532-42, 2012 Feb 01.
Article in English | MEDLINE | ID: mdl-22262180

ABSTRACT

BCL2L12 has been reported to be involved in post-mitochondrial apoptotic events in glioblastoma, but the role of BCL2L12A, a splicing variant of BCL2L12, remains unknown. In this study, we showed that BCL2L12 and BCL2L12A were overexpressed in glioblastoma multiforme (GBM). Large-scale yeast two-hybrid screening showed that BCL2L12 was a GSK3b binding partner in a testis cDNA library. Our data demonstrated that GSK3b interacts with BCL2L12 but not BCL2L12A, whose C terminus lacks a binding region. We found that a BCL2L12(153-191) fragment located outside of the C-terminal BH2 motif is responsible for GSK3b binding. In contrast, no interaction was detected between BCL2L12A and GSK3b. In vitro kinase and l-phosphatase assays showed that GSK3b phosphorylates BCL2L12 at S156, while this site is absent on BCL2L12A. Moreover, our data also showed that the BCL2L12(153-191) fragment directly interrupted GSK3bmediated Tau phosphorylation in a dose-dependent manner. Ectopic expression of GFP-fused BCL2L12 or BCL2L12A in U87MG cells leads to repression of apoptotic markers and protects against staurosporine (STS) insults, indicating an antiapoptotic role for both BCL2L12 and BCL2L12A. In contrast, no anti-apoptotic ability was seen in BCL2L12(S156A). When BCL2L12-expressing U87MG cells were co-administrated with STS and LiCl, cells underwent apoptosis. This effect could be reversed by LiCl. In short, we established a model to demonstrate that GSK3b interacts with and phosphorylates BCL2L12 and might also affect BCL2L12A to modulate the apoptosis signaling pathway in glioblastoma. These findings suggest that LiCl may be a prospective therapeutic agent against GBM.


Subject(s)
Apoptosis/drug effects , Brain Neoplasms/metabolism , Glioblastoma/metabolism , Glycogen Synthase Kinase 3/metabolism , Lithium Chloride/pharmacology , Muscle Proteins/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Signal Transduction , Brain Neoplasms/pathology , Cell Line, Tumor , Glioblastoma/pathology , Glycogen Synthase Kinase 3 beta , HEK293 Cells , Humans , Muscle Proteins/chemistry , Muscle Proteins/genetics , Phosphorylation , Protein Binding , Protein Isoforms/chemistry , Protein Isoforms/genetics , Protein Isoforms/metabolism , Protein Structure, Tertiary , Proto-Oncogene Proteins c-bcl-2/chemistry , Proto-Oncogene Proteins c-bcl-2/genetics , Signal Transduction/drug effects , Staurosporine/pharmacology , Two-Hybrid System Techniques , tau Proteins/metabolism
18.
Mol Med Rep ; 5(1): 118-25, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21971956

ABSTRACT

Cerebral vasospasm following subarachnoid hemorrhage (SAH) has been studied in terms of a contraction of the major cerebral arteries, but the effect of cerebrum tissue in SAH is not yet well understood. To gain insight into the biology of SAH-expressing cerebrum, we employed oligonucleotide microarrays to characterize the gene expression profiles of cerebrum tissue at the early stage of SAH. Functional gene expression in the cerebrum was analyzed 2 h following stage 1-hemorrhage in Sprague-Dawley rats. mRNA was investigated by performing microarray and quantitative real-time PCR analyses, and protein expression was determined by Western blot analysis. In this study, 18 upregulated and 18 downregulated genes displayed at least a 1.5-fold change. Five genes were verified by real-time PCR, including three upregulated genes [prostaglandin E synthase (PGES), CD14 antigen, and tissue inhibitor of metalloproteinase 1 (TIMP1)] as well as two downregulated genes [KRAB-zinc finger protein-2 (KZF-2) and γ-aminobutyric acid B receptor 1 (GABA B receptor)]. Notably, there were functional implications for the three upregulated genes involved in the inflammatory SAH process. However, the mechanisms leading to decreased KZF-2 and GABA B receptor expression in SAH have never been characterized. We conclude that oligonucleotide microarrays have the potential for use as a method to identify candidate genes associated with SAH and to provide novel investigational targets, including genes involved in the immune and inflammatory response. Furthermore, understanding the regulation of MMP9/TIMP1 during the early stages of SAH may elucidate the pathophysiological mechanisms in SAH rats.


Subject(s)
Cerebrum/immunology , Inflammation/genetics , Oligonucleotide Array Sequence Analysis , Subarachnoid Hemorrhage/genetics , Animals , Cerebrum/metabolism , Gene Expression Profiling , Inflammation/immunology , Intramolecular Oxidoreductases/genetics , Intramolecular Oxidoreductases/metabolism , Lipopolysaccharide Receptors/genetics , Lipopolysaccharide Receptors/metabolism , Male , Matrix Metalloproteinase 9/metabolism , Prostaglandin-E Synthases , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptors, GABA-B/genetics , Receptors, GABA-B/metabolism , Repressor Proteins/genetics , Repressor Proteins/metabolism , Subarachnoid Hemorrhage/immunology , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/metabolism
19.
Acta Neurochir (Wien) ; 153(11): 2211-7, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21833783

ABSTRACT

OBJECTIVE: Magnesium lithospermate B (MLB), a working extract from Salvia miltiorrhiza, was effective against coronary artery disease, ischemic stroke, and chronic renal disease. This study examined the effect of MLB on endothelin-1/endothelial nitric oxide synthase (eNOS) in a subarachnoid hemorrhage (SAH) animal model. METHODS: A rodent double-hemorrhage model was employed. Animals were randomly assigned to five groups (sham, SAH only, vehicle, 10 mg/kg/day MLB treatment, and pretreatment groups). A radiolabeled NOS Assay Kit was used to detect eNOS. Serum and cerebrospinal fluid sampling for ET-1 (ELISA) was measured. The basilar arteries (BAs) were garnered and sliced, and their cross-sectional areas were determined. In addition, NOS inhibitor nitro-arginine methyl ester (L-NAME) was employed in the SAH+ MLB treatment groups. RESULTS: Significant vasoconstriction was perceived in the SAH group (lumen patency: 44.6%, p < 0.01), but not in the MLB group (lumen patency: 89.3%). The ET-1 level was reduced in the MLP plus SAH group (34%, p < 0.01) when compared with the SAH groups (SAH only and vehicle). MLB dose-dependently increased the level of eNOS when compared with the vehicle plus SAH group. However, the administration of L-NAME reversed the expression of eNOS and vasoconstriction (lumen patency: 56.2%) in the MLB group. CONCLUSION: The enhanced expression of eNOS and decreased ET-1 levels in the MLB groups may reflect its anti-spastic effect. In the study of NOS, L-NAME reversed MLB's anti-vasospastic effect. This finding lends credence to the hypothesis that MLB modulates ET-1 levels through a NOS-dependent mechanism in the pathogenesis of cerebral vasospasm following SAH.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Endothelin-1/biosynthesis , Nitric Oxide/physiology , Subarachnoid Hemorrhage/physiopathology , Vasospasm, Intracranial/drug therapy , Animals , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Free Radical Scavengers/pharmacology , Free Radical Scavengers/therapeutic use , Male , Rats , Rats, Sprague-Dawley , Subarachnoid Hemorrhage/complications , Vasospasm, Intracranial/etiology , Vasospasm, Intracranial/physiopathology
20.
Pediatr Neurol ; 45(2): 135-7, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21763958

ABSTRACT

We describe a rare case of desmoplastic infantile ganglioglioma that appears to have transformed into a glioblastoma multiforme tumor in a 5-year-old girl. The patient was initially treated with total removal of the tumor, without subsequent radiation therapy or chemotherapy. She was in good health for 3 years, but manifested a sudden onset of seizures, followed by severe headache, vomiting, and left-sided weakness. Cranial magnetic resonance imaging revealed a large mass with rim enhancement at the previously operated site. The mass was completely removed, and the pathology report revealed glioblastoma multiforme. Postoperative chemotherapy and radiation treatment were administered. She has been followed for 11 years, and is alive without recurrence.


Subject(s)
Ganglioglioma/diagnosis , Glioblastoma/diagnosis , Neoplasms, Second Primary/diagnosis , Supratentorial Neoplasms/diagnosis , Adolescent , Cell Transformation, Neoplastic/pathology , Female , Ganglioglioma/pathology , Ganglioglioma/surgery , Glioblastoma/pathology , Glioblastoma/surgery , Humans , Neoplasms, Second Primary/pathology , Neoplasms, Second Primary/surgery , Supratentorial Neoplasms/pathology , Supratentorial Neoplasms/surgery
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