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1.
Dalton Trans ; 50(40): 14362-14373, 2021 Oct 19.
Article in English | MEDLINE | ID: mdl-34568879

ABSTRACT

The electrochemical water splitting by transition metal complexes is emerging very rapidly. The nickel complexes also play a very vital role in various biological activities. Here, three new ligands {H2mbhce = N'-(4-methyl-benzoyl), H2pchce = N'-(pyridine-carbonyl) and H2hbhce = N'-(2-hydroxy-benzoyl) hydrazine carbodithioic acid ethyl ester} and their corresponding Ni(II) complexes [Ni(Hmbhce)2(py)2] (1), [Ni(pchce)(o-phen)2]·CH3OH·H2O (2) and [Ni(hbhce)(o-phen)2]·1.75CHCl3·H2O (3) have been synthesized and fully characterized by various physicochemical and X-ray crystallography techniques. The photoluminescence study and thermal degradations were also examined. The treatment of K562 cells with the increasing concentrations of the nickel salts, ligands, and complexes 1, 2, and 3 showed dose-dependent cytotoxicity. The cytotoxic activity of ligands reveals that ligand H2mbhce is more potent in inhibiting the growth of tumor cells in comparison to other ligands H2pbhce and H2hbhce. Cytotoxicity assay results indicate that all complexes have remarkable cytotoxic potential in comparison to either nickel salts or the free ligands. Among these complexes, complex 1 has significantly better anti-tumor activity as compared to complexes 2 and 3. The electrochemical study of complexes 1, 2, and 3 for water oxidation reveals that all the complexes possess admirable electrocatalytic activity towards oxygen evolution reaction (OER) and have lower overpotential (328, 338, and 370 mV, respectively) than many previously reported complexes and RuO2 (390 mV). Among complexes 1, 2, and 3, complex-2 shows a better water oxidation response. Consequently, these complexes have great potential to be utilized in fuel cells. The more reliable electrochemical parameter TOF is also calculated for all three complexes.


Subject(s)
Antineoplastic Agents/pharmacology , Coordination Complexes/pharmacology , Electrochemical Techniques , Hydrazines/pharmacology , Nickel/pharmacology , Oxygen/chemistry , Thiocarbamates/pharmacology , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Cell Proliferation/drug effects , Coordination Complexes/chemical synthesis , Coordination Complexes/chemistry , Drug Screening Assays, Antitumor , Humans , Hydrazines/chemistry , K562 Cells , Molecular Structure , Nickel/chemistry , Thiocarbamates/chemistry
2.
J Neuroimmunol ; 299: 1-7, 2016 10 15.
Article in English | MEDLINE | ID: mdl-27725106

ABSTRACT

We evaluated the impact of latent toxoplasmosis (LT) on neurocognitive (NC) and neurobehavioural functioning in young adults with and without chronic HIV infection, using a standardised NC test battery, self-reported Beck Depression Inventory, Frontal System Behavior Scale, MINI-International Neuropsychiatric Interview and risk-assessment battery. 194 young adults (median age 24years, 48.2% males) with chronic HIV infection (HIV+) since childhood and 51 HIV seronegative (HIV-) participants were included. HIV+ individuals had good current immunological status (median CD4: 479 cells/µl) despite a low CD4 nadir (median: 93 cells/µl). LT (positive anti-Toxoplasma IgG antibodies) was present in one third of participants. The impairment rates in the HIV- with and without Toxo were not significantly different (p=0.17). However, we observed an increasing trend (p<0.001) in impairment rates with HIV and LT status: HIV-/LT- (6.1%); HIV-/LT+ (22%), HIV+/LT- (31%), HIV+/LT+ (49%). In a multivariable analysis using the entire study group there were main effects on cognition for HIV and also for LT. Within the HIV+ group LT was associated with worse performance globally (p=0.006), in memory (p=0.009), speed of information processing (p=0.01), verbal (p=0.02) and learning (p=0.02) domains. LT was not associated with depressive symptoms, frontal systems dysfunction or risk behaviors in any of the groups. HIV participants with lower Toxoplasma antibody concentration had worse NC performance, with higher GDS values (p=0.03) and worse learning (p=0.002), memory (p=0.006), speed of information processing (p=0.01) T scores. Latent Toxoplasmosis may contribute to NC impairment in young adults, including those with and without chronic HIV infection.


Subject(s)
Cognition Disorders/epidemiology , Cognition Disorders/psychology , HIV Infections/epidemiology , HIV Infections/psychology , Toxoplasmosis/epidemiology , Toxoplasmosis/psychology , Adult , Chronic Disease , Cognition Disorders/diagnosis , Cohort Studies , Cross-Sectional Studies , Female , HIV Infections/diagnosis , Humans , Male , Neuropsychological Tests , Toxoplasmosis/diagnosis , Young Adult
3.
Cell Oncol (Dordr) ; 34(6): 533-43, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21935712

ABSTRACT

BACKGROUND: Cervical cancer is a leading cause of cancer related deaths in women worldwide caused due to infection of high-risk human papillomaviruses. As JAK/STAT signalling pathway has been shown to play an important role during carcinogenesis, we studied the role of silencing of Suppressor of Cytokine Signalling-1 (SOCS-1) gene, a negative regulator of JAK/STAT pathway in cervical cancer. METHODS: The expression pattern of SOCS-1 mRNA and protein was analyzed in different stages of cervical tumor biopsies while normal cervical tissues served as controls. RT-PCR, immunohistochemistry and methylation-specific PCR (MSP) were performed to assess the expression pattern and promoter methylation of SOCS-1 gene in a total of 120 fresh surgically resected cervical tissue specimens comprising precancer (n = 12), cancer (n = 78) and normal controls (n = 30). RESULTS: Compared with expression of SOCS-1 in normal tissues, 64% of the tumor tissues expressed either undetectable or reduced expression. Aberrant promoter methylation of SOCS-1 was found in 61% of the cervical tumor tissues. SOCS-1 expression and methylation were significantly associated with severity of the disease (p < 0.01). CONCLUSION: We demonstrate for the first time the transcriptional inactivation of SOCS-1 gene due to hypermethylation and synergism with HPV infection which may play an important role in cervical carcinoma.


Subject(s)
DNA Methylation/genetics , Promoter Regions, Genetic/genetics , Suppressor of Cytokine Signaling Proteins/genetics , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/metabolism , Female , Genetic Predisposition to Disease , Humans , In Vitro Techniques , Papillomaviridae/genetics , Papillomaviridae/isolation & purification , Suppressor of Cytokine Signaling 1 Protein , Suppressor of Cytokine Signaling Proteins/metabolism , Uterine Cervical Neoplasms/virology
4.
J Forensic Odontostomatol ; 29(1): 51-6, 2011 Jul 01.
Article in English | MEDLINE | ID: mdl-21841269

ABSTRACT

Teeth are considered as a useful adjunct for sex assessment and may play an important role in constructing a post-mortem profile. The Dimodent method is based on the high degree of sex discrimination obtained with the mandibular canine and the high correlation coefficients between mandibular canine and lateral incisor mesiodistal (MD) and buccolingual (BL) dimensions. This has been evaluated in the French and Lebanese, but no study exists on its efficacy in Indians. Here, we have applied the 'Dimodent' equation on an Indian sample (100 males, 100 females; age range of 19-27yrs). Additionally, a population-specific Dimodent equation was derived using logistic regression analysis and applied to our sample. Also, the sex determination potential of MD and BL measurements of mandibular lateral incisors and canines, individually, was assessed. We found a poor sex assessment accuracy using the Dimodent equation of Fronty (34.5%) in our Indian sample, but the populationspecific Dimodent equation gave a better accuracy (72%).Thus, it appears that sexual dimorphism in teeth is population-specific; consequently the Dimodent equation has to be derived individually in different populations for use in sex assessment. The mesiodistal measurement of the mandibular canine alone gave a marginally higher accuracy (72.5%); therefore, we suggest the use of mandibular canines alone rather than the Dimodent method.


Subject(s)
Cuspid/anatomy & histology , Forensic Dentistry/methods , Models, Statistical , Sex Determination Analysis , Adult , Autopsy , Female , Humans , Incisor/anatomy & histology , India , Male , Mandible/anatomy & histology , Mouth/anatomy & histology , Reference Values , Young Adult
5.
J Laryngol Otol ; 125(4): 343-7, 2011 Apr.
Article in English | MEDLINE | ID: mdl-20955634

ABSTRACT

INTRODUCTION: Vestibular evoked myogenic potentials are short latency electrical impulses that are produced in response to higher level acoustic stimuli. They are used clinically to diagnose sacculocollic pathway dysfunction. AIM: This study aimed to compare the vestibular evoked myogenic potential responses elicited by click stimuli and short duration tone burst stimuli, in normal hearing individuals. METHOD: Seventeen subjects participated. In all subjects, we assessed vestibular evoked myogenic potentials elicited by click and short duration tone burst stimuli. RESULTS AND CONCLUSION: The latency of the vestibular evoked myogenic potential responses (i.e. the p13 and n23 peaks) was longer for tone burst stimuli compared with click stimuli. The amplitude of the p13-n23 waveform was greater for tone burst stimuli than click stimuli. Thus, the click stimulus may be preferable for clinical assessment and identification of abnormalities as this stimulus has less variability, while a low frequency tone burst stimulus may be preferable when assessing the presence or absence of vestibular evoked myogenic potential responses.


Subject(s)
Acoustic Stimulation/methods , Vestibular Evoked Myogenic Potentials/physiology , Acoustic Stimulation/instrumentation , Adult , Female , Humans , Male , Saccule and Utricle/physiology , Time Factors , Vestibular Function Tests/instrumentation , Vestibular Function Tests/methods , Vestibular Nerve/physiology , Young Adult
6.
Eur Respir J ; 36(4): 885-92, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20185420

ABSTRACT

Assessment of total plasma DNA can be a promising noninvasive tool for monitoring the effect of cytotoxic therapies in order to predict therapeutic efficacy at an early stage. Cell-free plasma DNA levels were quantified before the first, second and third cycle of chemotherapy in 42 patients with advanced nonsmall cell lung cancer and correlated with response to therapy, as assessed by computed tomography following the third chemotherapy cycle. A significantly lower plasma DNA level, measured before various treatment cycles, was found in patients with remission or stable disease than in those with progression. Higher levels and insufficient decrease in plasma DNA levels during the course of chemotherapy indicated poor outcome. For predicting insufficient therapy response, a sensitivity of 26.9% was achieved at 100% specificity using plasma DNA levels before the first therapy cycle. Prediction of disease progression was achieved with a sensitivity of 35.7% at 100% specificity using plasma DNA levels before the first therapy cycle. Monitoring of plasma DNA levels during the course of chemotherapy could identify patients who are likely to exhibit an insufficient therapeutic response and disease progression at an early stage. This may help in individualising treatment, and could lead to better management of advanced-stage lung cancer.


Subject(s)
Carcinoma, Non-Small-Cell Lung/blood , Carcinoma, Non-Small-Cell Lung/genetics , DNA/blood , Lung Neoplasms/blood , Lung Neoplasms/genetics , Adult , Aged , Aged, 80 and over , Antineoplastic Agents/pharmacology , Biomarkers/metabolism , Biopsy , Carcinoma, Non-Small-Cell Lung/drug therapy , DNA/genetics , Humans , Lung Neoplasms/drug therapy , Medical Oncology/methods , Middle Aged , Prospective Studies , Sensitivity and Specificity
7.
Mol Cell Biochem ; 330(1-2): 193-9, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19421717

ABSTRACT

The constitutively activated STAT family members, particularly STAT3, have been shown to possess transforming properties, and are strongly correlated with tumor development and progression. STAT3 transmits signals from many cytokines and growth factors to target genes in the nucleus through the Jak/Stat signaling pathway. HPV is the main etiological factor in the development of cervical cancer. In the current study, the expression of STAT3 was analyzed in various stages of HPV-mediated cervical carcinogenesis. Tissue biopsies from 100 patients with cervical cancer of different stages and normal tissues from patients undergoing hysterectomy were selected for studying the HPV status and STAT3 expression. HPV status of each corresponding biopsy was analyzed by PCR and typing. The mRNA expression was analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR). HPV infection was detected in majority of cases: 75% (9/12) in precancer, 85% (34/40) stage I & II, and 95% (36/38) in stage III & IV of cervical cancer cases by L1 PCR. Further sub typing revealed HPV16 in 100% (9/9) of L1 positives in precancerous & 90% (63/70) in different stages of cancer. Significant level of STAT3 mRNA expression was predominantly found in cervical cancer cases as compared to normal controls (P = 0.001). We also found a significant correlation of STAT3 expression in cases infected with HPV (P = 0.001). Our results indicate a potentially interactive effect between HPV 16/18 and transcriptional activation of STAT3 gene in cervical carcinogenesis. To our knowledge, this is the first such study to be reported from India. Further investigations are needed to determine the influence of STAT3 expression on cervical carcinogenesis and its possible interaction with HPV infection status.


Subject(s)
Gene Expression Regulation, Neoplastic , Papillomaviridae/isolation & purification , STAT3 Transcription Factor/genetics , Uterine Cervical Neoplasms/etiology , Case-Control Studies , Female , Human papillomavirus 16/isolation & purification , Human papillomavirus 18/isolation & purification , Humans , India/epidemiology , Neoplasm Staging , Papillomavirus Infections/complications , Papillomavirus Infections/diagnosis , RNA, Messenger/analysis , Transcriptional Activation , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/virology
8.
Oncogene ; 27(13): 1886-93, 2008 Mar 20.
Article in English | MEDLINE | ID: mdl-17922035

ABSTRACT

Heat shock factor 1 (HSF1), the transcriptional activator of the heat shock genes, is increasingly implicated in cancer. We have shown that HSF1 binds to the corepressor metastasis-associated protein 1 (MTA1) in vitro and in human breast carcinoma samples. HSF1-MTA1 complex formation was strongly induced by the transforming ligand heregulin and complexes incorporated a number of additional proteins including histone deacetylases (HDAC1 and 2) and Mi2alpha, all components of the NuRD corepressor complex. These complexes were induced to assemble on the chromatin of MCF7 breast carcinoma cells and associated with the promoters of estrogen-responsive genes. Such HSF1 complexes participate in repression of estrogen-dependent transcription in breast carcinoma cells treated with heregulin and this effect was inhibited by MTA1 knockdown. Repression of estrogen-dependent transcription may contribute to the role of HSF1 in cancer.


Subject(s)
Breast Neoplasms/genetics , DNA-Binding Proteins/physiology , Estrogens/pharmacology , Histone Deacetylases/metabolism , Repressor Proteins/metabolism , Transcription Factors/physiology , Transcription, Genetic , Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/metabolism , Autoantigens/genetics , Autoantigens/metabolism , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Chromatin/metabolism , Chromatin Immunoprecipitation , DNA Helicases/genetics , DNA Helicases/metabolism , Gene Expression Regulation, Neoplastic , Heat Shock Transcription Factors , Histone Deacetylase 1 , Histone Deacetylase 2 , Histone Deacetylase Inhibitors , Histone Deacetylases/genetics , Humans , Immunoenzyme Techniques , Mi-2 Nucleosome Remodeling and Deacetylase Complex , Neuregulin-1/pharmacology , Promoter Regions, Genetic , Repressor Proteins/antagonists & inhibitors , Repressor Proteins/genetics , Trans-Activators , Tumor Cells, Cultured
9.
Oncogene ; 25(1): 20-31, 2006 Jan 05.
Article in English | MEDLINE | ID: mdl-16158055

ABSTRACT

The MUC1 heterodimeric transmembrane glycoprotein is aberrantly overexpressed by most human carcinomas. The MUC1 C-terminal subunit localizes to mitochondria and blocks stress-induced activation of the intrinsic apoptotic pathway. How MUC1 is delivered to mitochondria is not known. The present studies demonstrate that MUC1 forms intracellular complexes with HSP70 and HSP90. We show that the MUC1 cytoplasmic domain binds directly to HSP70 in vitro. By contrast, binding of MUC1 to HSP90 in vitro is induced by c-Src-mediated phosphorylation of the MUC1 cytoplasmic domain. c-Src also increases binding of MUC1 to HSP90 in cells. In concert with these results, we show that heregulin (HRG), a ligand for ErbB receptors, activates c-Src and, in turn, stimulates binding of MUC1 to HSP90. We also show that inhibitors of c-Src or HSP90 block HRG-induced targeting of MUC1 to mitochondria and integration of MUC1 into the mitochondrial outer membrane. These findings indicate that MUC1 is delivered to mitochondria by a mechanism involving activation of the ErbB receptor-->c-Src pathway and transport by the molecular chaperone HSP70/HSP90 complex.


Subject(s)
Antigens/physiology , Glycoproteins/physiology , HSP90 Heat-Shock Proteins/metabolism , Mitochondria/metabolism , Molecular Chaperones/chemistry , Mucins/physiology , Neuregulin-1/chemistry , src-Family Kinases/metabolism , Amino Acid Sequence , Animals , Antigens/metabolism , Antigens, Neoplasm , Cell Line , Cell Line, Tumor , Cell Membrane/metabolism , Cytoplasm/metabolism , Cytosol/metabolism , Electrophoresis, Polyacrylamide Gel , ErbB Receptors/metabolism , Glycoproteins/metabolism , HSP70 Heat-Shock Proteins/metabolism , HeLa Cells , Humans , Immunoblotting , Immunoglobulin G/chemistry , Immunoprecipitation , Mass Spectrometry , Mice , Models, Biological , Molecular Chaperones/metabolism , Molecular Sequence Data , Mucin-1 , Mucins/metabolism , Neuregulin-1/metabolism , Phosphorylation , Protein Binding , Protein Structure, Tertiary , Signal Transduction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Subcellular Fractions/metabolism , Time Factors , Transfection
10.
Apoptosis ; 9(6): 677-90, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15505411

ABSTRACT

Many members of tumor necrosis factor (TNF) superfamily are characterized by their ability to induce apoptosis once they bind in a homotrimeric manner to their cognate receptors. The receptor activator of nuclear factor-kappaB ligand (RANKL), a member of the TNF superfamily identified seven years ago, was originally described as a factor that induced osteoclastogenesis and dendritic cell survival. Recent observations indicate that a growth inhibitory and apoptosis-inducing activity is associated with RANKL, as is the case for other members of TNF superfamily. This review describes the possible mechanisms of induction of RANKL-induced growth inhibition/apoptosis and discusses the role of various components in RANKL-signaling in this phenomenon, including TNF receptor-associated factor (TRAF)-6, nuclear factor-kappaB (NF-kappaB), c-jun N-terminal kinase JNK), phosphatidylinositol-3 kinase (PI3K).


Subject(s)
Apoptosis , Carrier Proteins/metabolism , Membrane Glycoproteins/metabolism , Animals , Humans , JNK Mitogen-Activated Protein Kinases/metabolism , Models, Biological , NF-kappa B/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins/metabolism , Proto-Oncogene Proteins c-akt , RANK Ligand , Receptor Activator of Nuclear Factor-kappa B , TNF Receptor-Associated Factor 6/metabolism
11.
Cell Growth Differ ; 12(9): 465-70, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11571229

ABSTRACT

The cellular response to oxidative stress includes the release of mitochondrial cytochrome c and the induction of apoptosis. Here we show that treatment of diverse cells with hydrogen peroxide (H2O2) induces the targeting of protein kinase C delta (PKCdelta) to mitochondria. The results demonstrate that H2O2-induced activation of PKCdelta is necessary for translocation of PKCdelta from the cytoplasm to the mitochondria. The results also show that mitochondrial targeting of PKCdelta is associated with the loss of mitochondrial transmembrane potential and release of cytochrome c. The functional importance of this event is also supported by the demonstration that H2O2-induced apoptosis is blocked by the inhibition of PKCdelta activation and translocation to mitochondria. These findings indicate that mitochondrial targeting of PKCdelta is required, at least in part, for the apoptotic response of cells to oxidative stress.


Subject(s)
Isoenzymes/metabolism , Mitochondria/metabolism , Oxidative Stress/physiology , Protein Kinase C/metabolism , 3T3 Cells , Animals , Cytochrome c Group/metabolism , Enzyme Activation , Humans , Hydrogen Peroxide/pharmacology , Mice , Oxidants/pharmacology , Protein Kinase C-delta , Protein Transport , Proto-Oncogene Proteins c-abl/metabolism , Reactive Oxygen Species/metabolism , Tumor Cells, Cultured , U937 Cells
12.
J Biol Chem ; 276(20): 17281-5, 2001 May 18.
Article in English | MEDLINE | ID: mdl-11350980

ABSTRACT

The ubiquitously expressed c-Abl tyrosine kinase is activated in the response of cells to genotoxic and oxidative stress. The present study demonstrates that reactive oxygen species (ROS) induce targeting of c-Abl to mitochondria. We show that ROS-induced localization of c-Abl to mitochondria is dependent on activation of protein kinase C (PKC)delta and the c-Abl kinase function. Targeting of c-Abl to mitochondria is associated with ROS-induced loss of mitochondrial transmembrane potential. The results also demonstrate that c-Abl is necessary for ROS-induced depletion of ATP and the activation of a necrosis-like cell death. These findings indicate that the c-Abl kinase targets to mitochondria in response to oxidative stress and thereby mediates mitochondrial dysfunction and cell death.


Subject(s)
Cell Death/physiology , Hydrogen Peroxide/toxicity , Mitochondria/physiology , Oxidative Stress/physiology , Proto-Oncogene Proteins c-abl/metabolism , Animals , Cell Death/drug effects , Genes, abl/drug effects , Humans , Intracellular Membranes/physiology , Isoenzymes/metabolism , Membrane Potentials , Mice , Mice, Knockout , Necrosis , Protein Kinase C/metabolism , Protein Kinase C-delta , Protein-Tyrosine Kinases/metabolism , Reactive Oxygen Species/physiology , Recombinant Fusion Proteins/metabolism , Transfection , Tumor Cells, Cultured , U937 Cells
13.
Int Immunopharmacol ; 1(1): 155-65, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11367513

ABSTRACT

Previously, we have observed a modulation in the bone marrow hematopoiesis and alteration in the repertoire of blood monocytes and lymphocytes in mice bearing a spontaneous T cell lymphoma, designated as Dalton's lymphoma (DL). In the present investigation, we show that in vivo or in vitro treatment of bone marrow cells (BMC) with gangliosides of DL (DLG) results in inhibition of proliferative ability and alteration of colony-forming ability (CFA) of BMC. BMC incubated with DLG also showed a decrease in cell viability in a concentration-dependent manner. BMC colony-forming assays in the presence of macrophage-colony-stimulating factor (M-CSF) showed a dose-dependent decrease in the number of colonies with a concomitant decrease in macrophage- and granulocyte macrophage-colony-forming units (CFU-M and CFU-GM, respectively) and an increase in granulocyte-CFU (CFU-G). Neuraminidase treatment of DLG abrogated their inhibitory action on BMC. Further, antibodies to GD3 and to lesser extent GM2 neutralized the inhibitory effect of DLG on BMC.


Subject(s)
Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Gangliosides/pharmacology , Animals , Cell Differentiation/drug effects , Cell Division/drug effects , Colony-Forming Units Assay , Female , Gangliosides/isolation & purification , Immunosuppressive Agents/isolation & purification , Immunosuppressive Agents/pharmacology , In Vitro Techniques , Lymphoma, T-Cell/chemistry , Lymphoma, T-Cell/immunology , Male , Mice , Mice, Inbred BALB C , Neuraminidase
14.
Mol Cell Biol ; 20(14): 4979-89, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10866655

ABSTRACT

The c-Abl protein tyrosine kinase is activated by certain DNA-damaging agents and regulates induction of the stress-activated c-Jun N-terminal protein kinase (SAPK). Here we show that nuclear c-Abl associates with MEK kinase 1 (MEKK-1), an upstream effector of the SEK1-->SAPK pathway, in the response of cells to genotoxic stress. The results demonstrate that the nuclear c-Abl binds to MEKK-1 and that c-Abl phosphorylates MEKK-1 in vitro and in vivo. Transient-transfection studies with wild-type and kinase-inactive c-Abl demonstrate c-Abl kinase-dependent activation of MEKK-1. Moreover, c-Abl activates MEKK-1 in vitro and in response to DNA damage. The results also demonstrate that c-Abl induces MEKK-1-mediated phosphorylation and activation of SEK1-SAPK in coupled kinase assays. These findings indicate that c-Abl functions upstream of MEKK-1-dependent activation of SAPK in the response to genotoxic stress.


Subject(s)
DNA Damage/physiology , MAP Kinase Kinase 4 , MAP Kinase Kinase Kinase 1 , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins c-abl/metabolism , Animals , Cells, Cultured , Cisplatin/pharmacology , DNA Damage/radiation effects , Enzyme Activation/radiation effects , Humans , JNK Mitogen-Activated Protein Kinases , Mice , Mice, Mutant Strains , Mitogen-Activated Protein Kinase Kinases/genetics , Mitogen-Activated Protein Kinase Kinases/metabolism , Mitogen-Activated Protein Kinases/metabolism , Protein Serine-Threonine Kinases/genetics , Proto-Oncogene Proteins c-abl/drug effects , Proto-Oncogene Proteins c-abl/genetics , Radiation, Ionizing , U937 Cells
15.
Immunol Lett ; 72(1): 39-48, 2000 Apr 03.
Article in English | MEDLINE | ID: mdl-10789680

ABSTRACT

The present study was conducted to investigate the mode of cell death of bone marrow cells (BMC) treated in vivo or in vitro with gangliosides (DLG) derived from a T cell lymphoma, designated as Dalton's lymphoma (DL). BMC undergoing cell death showed morphological features characteristic to apoptosis such as cell shrinkage, plasma membrane blebbing and densely stained chromatin. DLG-induced apoptosis was further confirmed by analysis of DNA from DLG-treated BMC by flow cytometry and agarose gel electrophoresis which showed an increase in hypodiploid DNA and internucleosomal DNA cleavage, respectively. Immunoblotting of p53 and Bax demonstrated an increase in the levels of p53 and Bax proteins. BMC treated with DLG also showed translocation of NF-kappaB protein into the nucleus. Antibodies to ganglioside G(D3), a constituent of DLG, neutralized the effect of DLG. Further, supplementation of DLG-containing BMC cultures with granulocyte macrophage-colony stimulating factor (GM-CSF) or macrophage-colony stimulating factor (M-CSF) prevented the apoptosis-inducing action of DLG.


Subject(s)
Apoptosis , Bone Marrow Cells/cytology , Gangliosides/metabolism , Gangliosides/pharmacology , Lymphoma, T-Cell/metabolism , Proto-Oncogene Proteins c-bcl-2 , Animals , Cell Survival/drug effects , Female , Germ-Free Life , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Hematopoiesis , Macrophage Colony-Stimulating Factor/pharmacology , Male , Mice , Mice, Inbred BALB C , NF-kappa B/metabolism , Proto-Oncogene Proteins/metabolism , Tumor Cells, Cultured , Tumor Suppressor Protein p53/metabolism , bcl-2-Associated X Protein
16.
Oncogene ; 19(16): 1975-81, 2000 Apr 13.
Article in English | MEDLINE | ID: mdl-10803458

ABSTRACT

The release of mitochondrial cytochrome c by genotoxic stress induces the formation of a cytosolic complex with Apaf-1 (mammalian CED4 homolog) and thereby the activation of procaspase-3 (cas-3) and procaspase-9 (cas-9). Here we demonstrate that heat-shock protein 27 (Hsp27) inhibits cytochrome c (cyt c)-dependent activation of cas-3. Hsp27 had no effect on cyt c release, Apaf-1 and cas-9 activation. By contrast, our results show that Hsp27 associates with cas-3, but not Apaf-1 or cas-9, and inhibits activation of cas-3 by cas-9-mediated proteolysis. Furthermore, the present results demonstrate that immunodepletion of Hsp27 depletes cas-3. Importantly, treatment of cells with DNA damaging agents dissociates the Hsp27/cas-3 complex and relieves inhibition of cas-3 activation. These findings define a novel function for Hsp27 and provide the first evidence that a heat shock protein represses cas-3 activation.


Subject(s)
Caspases/metabolism , Cytochrome c Group/metabolism , Enzyme Precursors/metabolism , Heat-Shock Proteins/metabolism , Actins/metabolism , Apoptotic Protease-Activating Factor 1 , Caspase 3 , Caspase 9 , Caspases/drug effects , Caspases/immunology , Cell Line/drug effects , Cell Line/radiation effects , Cell-Free System , Cytarabine/pharmacology , Cytochrome c Group/drug effects , Cytochrome c Group/radiation effects , Cytosol/metabolism , Enzyme Activation , Enzyme Inhibitors/pharmacology , Enzyme Precursors/drug effects , Enzyme Precursors/immunology , Heat-Shock Proteins/genetics , Heat-Shock Proteins/immunology , Heat-Shock Proteins/radiation effects , Humans , Immunoblotting , Isoenzymes/metabolism , Methyl Methanesulfonate/pharmacology , Oligopeptides/metabolism , Protein Kinase C/metabolism , Protein Kinase C-delta , Proteins/metabolism , Staurosporine/pharmacology
17.
EMBO J ; 19(5): 1087-97, 2000 Mar 01.
Article in English | MEDLINE | ID: mdl-10698949

ABSTRACT

Hormones and growth factors induce protein translation in part by phosphorylation of the eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4E-BP1). The rapamycin and FK506-binding protein (FKBP)-target 1 (RAFT1, also known as FRAP) is a mammalian homolog of the Saccharomyces cerevisiae target of rapamycin proteins (mTOR) that regulates 4E-BP1. However, the molecular mechanisms involved in growth factor-initiated phosphorylation of 4E-BP1 are not well understood. Here we demonstrate that protein kinase Cdelta (PKCdelta) associates with RAFT1 and that PKCdelta is required for the phosphorylation and inactivation of 4E-BP1. PKCdelta-mediated phosphorylation of 4E-BP1 is wortmannin resistant but rapamycin sensitive. As shown for serum, phosphorylation of 4E-BP1 by PKCdelta inhibits the interaction between 4E-BP1 and eIF4E and stimulates cap-dependent translation. Moreover, a dominant-negative mutant of PKCdelta inhibits serum-induced phosphorylation of 4E-BP1. These findings demonstrate that PKCdelta associates with RAFT1 and thereby regulates phosphorylation of 4E-BP1 and cap-dependent initiation of protein translation.


Subject(s)
Carrier Proteins/genetics , Isoenzymes/genetics , Phosphoproteins/genetics , Phosphotransferases (Alcohol Group Acceptor) , Protein Biosynthesis , Protein Kinase C/genetics , Androstadienes/pharmacology , Animals , Anti-Bacterial Agents/pharmacology , Carrier Proteins/metabolism , Cell Line , Isoenzymes/metabolism , Peptide Initiation Factors/genetics , Peptide Initiation Factors/metabolism , Phosphoproteins/metabolism , Phosphorylation , Protein Kinase C/metabolism , Protein Kinase C-delta , RNA, Messenger/genetics , Sirolimus/metabolism , Sirolimus/pharmacology , TOR Serine-Threonine Kinases , Wortmannin
18.
J Biol Chem ; 274(43): 30657-63, 1999 Oct 22.
Article in English | MEDLINE | ID: mdl-10521452

ABSTRACT

Related adhesion focal tyrosine kinase (RAFTK) (also known as PYK2) is a cytoplasmic tyrosine kinase related to the focal adhesion kinase (FAK) p125(FAK). RAFTK is rapidly phosphorylated on tyrosine residues in response to various stimuli, such as tumor necrosis factor-alpha, changes in osmolarity, elevation in intracellular calcium concentration, lysophosphatidic acid, and bradykinin. Overexpression of RAFTK induces activation of c-Jun amino-terminal kinase (also known as stress-activated protein kinase), mitogen-activated protein kinase (MAPK), and p38 MAPK. The present studies demonstrate that RAFTK binds constitutively to the protein tyrosine phosphatase SHPTP1. In contrast to PTP1B, overexpression of wild-type SHPTP1 blocks tyrosine phosphorylation of RAFTK. The results further demonstrate that RAFTK is a direct substrate of SHPTP1 in vitro. Moreover, treatment of PC12 cells with bradykinin is associated with inhibition in tyrosine phosphorylation of RAFTK in the presence of SHPTP1. Furthermore, in contrast to the phosphatase-dead SHPTP1 C453S mutant, overexpression of wild-type SHPTP1 blocks interaction of RAFTK with the SH2-domain of c-Src and inhibits RAFTK-mediated MAPK activation. Significantly, cotransfection of RAFTK with SHPTP1 did not inhibit RAFTK-mediated c-Jun amino-terminal kinase activation. Taken together, these findings suggest that SHPTP1 plays a negative role in PYK2/RAFTK signaling by dephosphorylating RAFTK.


Subject(s)
Cell Adhesion Molecules/metabolism , Protein Tyrosine Phosphatases/metabolism , Protein-Tyrosine Kinases/metabolism , Signal Transduction/physiology , Cell Line , Focal Adhesion Kinase 1 , Focal Adhesion Kinase 2 , Focal Adhesion Protein-Tyrosine Kinases , Homeostasis , Humans , Intracellular Signaling Peptides and Proteins , Kinetics , Mutagenesis, Site-Directed , Oligopeptides , Peptides , Protein Tyrosine Phosphatase, Non-Receptor Type 11 , Protein Tyrosine Phosphatase, Non-Receptor Type 6 , Recombinant Proteins/metabolism , SH2 Domain-Containing Protein Tyrosine Phosphatases , Transfection , U937 Cells , src Homology Domains
19.
Blood ; 93(5): 1482-6, 1999 Mar 01.
Article in English | MEDLINE | ID: mdl-10029574

ABSTRACT

Whether Kaposi's sarcoma herpesvirus (KSHV) is associated with multiple myeloma (MM) remains controversial. We assayed for KSHV DNA sequences in long-term bone marrow stromal cells (BMSCs) from 26 patients with MM and 4 normal donors. Polymerase chain reaction (PCR) using primers which amplify a KSHV gene sequence to yield a 233-bp fragment (KS330233 within open reading frame 26) was negative in all cases. Aliquots of these PCR products were used as templates in subsequent nested PCR, with primers that amplify a 186-bp product internal to KS330233. BMSCs from 24 of 26 (92%) patients with MM and 1 of 4 normal donors were KSHV PCR+. DNA sequence analyses showed interpatient specific mutations (2 to 3 bp). Both Southern blot and sequence analyses confirmed the specificity of PCR results. The presence of the KSHV gene sequences was further confirmed by amplifying T 1.1 (open reading frame [ORF] K7) and viral cyclin D (ORF 72), two other domains within the KSHV genome. Immunohistochemical studies of KSHV PCR+ MM BMSCs demonstrate expression of dendritic cell (DC) lineage markers (CD68, CD83, and fascin). Serological studies for the presence of KSHV lytic or latent antibodies were performed using sera from 53 MM patients, 12 normal donors, and 5 human immunodeficiency virus (HIV)/KSHV+ patients. No lytic or latent antibodies were present in sera from either MM patients or normal donors. Taken together, these findings show that KSHV DNA sequences are detectable in BMSCs from the majority of MM patients, but that serologic responses to KSHV are not present. Ongoing studies are defining whether the lack of antibody response is caused by the absence of ongoing infection, the presence of a novel viral strain associated with MM, or underlying immunodeficiency in these patients.


Subject(s)
Bone Marrow Cells/virology , Herpesvirus 8, Human/isolation & purification , Multiple Myeloma/virology , Stromal Cells/virology , Bone Marrow Cells/pathology , Cyclin D , Cyclins/genetics , DNA, Viral/analysis , Herpesvirus 8, Human/genetics , Humans , Multiple Myeloma/pathology , Stromal Cells/pathology , Viral Proteins/analysis , Viral Proteins/genetics
20.
Mol Cell Biol ; 18(12): 7216-24, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9819408

ABSTRACT

The DF3/MUC1 mucin-like glycoprotein is highly overexpressed in human carcinomas. Recent studies have demonstrated that the cytoplasmic domain of MUC1 interacts with beta-catenin. Here we show that MUC1 associates with glycogen synthase kinase 3beta (GSK3beta). GSK3beta binds directly to an STDRSPYE site in MUC1 and phosphorylates the serine adjacent to proline. Phosphorylation of MUC1 by GSK3beta decreases binding of MUC1 to beta-catenin in vitro and in vivo. GSK3beta-mediated phosphorylation of MUC1 had no apparent effect on beta-catenin levels or the transcriptional coactivation function of beta-catenin. The results, however, demonstrate that MUC1 expression decreases binding of beta-catenin to the E-cadherin cell adhesion molecule. Negative regulation of the beta-catenin-MUC1 interaction by GSK3beta is associated with restoration of the complex between beta-catenin and E-cadherin. These findings indicate that GSK3beta decreases the interaction of MUC1 with beta-catenin and that overexpression of MUC1 in the absence of GSK3beta activity inhibits formation of the E-cadherin-beta-catenin complex.


Subject(s)
Antigens, Neoplasm/metabolism , Breast Neoplasms/enzymology , Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Cytoskeletal Proteins/metabolism , Mucin-1/metabolism , Trans-Activators , Amino Acid Sequence , Binding Sites , Binding, Competitive , Cadherins/metabolism , Down-Regulation/physiology , Glycogen Synthase Kinase 3 , Glycogen Synthase Kinases , Humans , Molecular Sequence Data , Mutagenesis, Site-Directed/genetics , Phosphorylation , Recombinant Fusion Proteins/genetics , Transfection/genetics , Tumor Cells, Cultured , beta Catenin
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