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OBJECTIVE: To investigate the survival ability of Staphylococcus epidermidis (S. epidermidis) wildtype (WT), saeRS mutant (SAE) and saeRS complementary (SAEC) strains under different concentrations of glucose. METHODS: We measured the survival ability, biofilm forming ability, medium acidity and antimicrobial susceptibility of S. epidermidis in medium containing different concentrations of glucose. RESULTS: Compared with WT, the survival ability, biofilm forming ability and resistance to antibiotics (such as penicillin, oxacillin, gentamicin, ciprofloxacin and amikacin) of saeRS mutant increased significantly in response to glucose. SAE and SAEC showed the strongest survival ability and biofilm forming ability when grown in medium containing 14 mmol/L glucose and 28 mmol/L, respectively. WT showed no significant different survival and biofilm forming abilities when cultured with various concentrations of glucose. The medium acidity of saeRS mutant (pH=8.07) was lower than the WT (pH=7.0) in the presence of 14 mmol/L glucose. CONCLUSION: SaeRS may influence the survival ability of S. epidermidis by affecting glucose utilization.
Assuntos
Biofilmes , Transdução de Sinais , Staphylococcus epidermidis/crescimento & desenvolvimento , Antibacterianos/farmacologia , Meios de Cultura , Glucose/metabolismo , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/genéticaRESUMO
The aim of this study was to identify novel biomarkers for the diagnosis of, and potential therapeutic targets for, hepatocellular carcinoma (HCC). Multilectin affinity chromatography was used to enrich N-linked glycoproteins from nontumorous liver and HCC tissues followed by 2DE and protein identification by MS. Twenty-eight differentially expressed proteins were identified. Western blotting validated consistently lower concentrations of human liver carboxylesterase 1 and haptoglobin, and higher concentration of procathepsin D (pCD) in HCC tissues. Knockdown of cathepsin D (CD) expression mediated by siRNA significantly inhibited the in vitro invasion of two HCC cell lines, SNU449 and SNU473, which normally secrete high-levels of CD. Prefractionation using individual lectins demonstrated an elevation in ConA-binding glycoforms of proCD and CD in HCC tissues. In the serum of HCC patients, "ConA-binding proCD" (ConA-pCD) is significantly increased in concentration and this increase is comprised of several distinct upregulated acidic isoforms (pI 4.5-5.5). Receiver operating characteristic analysis showed that the sensitivity and specificity of serum ConA-pCD for HCC diagnosis were 85% and 80%, respectively. This is the first report that serum ConA-pCD is increased significantly in HCC and is potentially useful as a serological biomarker for diagnosis of HCC.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/sangue , Catepsina D/sangue , Precursores Enzimáticos/sangue , Neoplasias Hepáticas/sangue , Proteômica/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Western Blotting , Hidrolases de Éster Carboxílico/análise , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Catepsina D/análise , Catepsina D/genética , Catepsina D/metabolismo , Linhagem Celular Tumoral , Concanavalina A/metabolismo , Precursores Enzimáticos/análise , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Glicoproteínas/análise , Glicoproteínas/sangue , Glicoproteínas/metabolismo , Haptoglobinas/análise , Humanos , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Interferência de RNA , RNA Interferente Pequeno/genéticaRESUMO
Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Cisplatino/farmacologia , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Neoplasias Esofágicas/metabolismo , Proteoma/metabolismo , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Neoplasias Esofágicas/patologia , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Oxirredutases Intramoleculares/metabolismo , Marcação por Isótopo , Fatores Inibidores da Migração de Macrófagos/metabolismo , Proteômica , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Tiorredoxinas/metabolismoRESUMO
The title compound, {[Tm(C(7)H(3)NO(4))(C(7)H(4)NO(4))(H(2)O)(2)]·2H(2)O}(n), is isotypic with the analogous Tb(III) compound [Li et al. (2009 â¶). Acta Cryst. E65, m410]. All interatomic distances and angles and the hydrogen-bond geometries are very similar for the two structures. The refined Flack parameter of 0.49â (2) suggests inversion twinning.
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Esophageal cancer (EC) persists to be a leading cancer-related death in northern China. Clinical outcome of EC is the most dismal among many types of digestive tumors because EC at early stage is asymptomatic. The current study used 2-DE-based proteomics to identify differentially expressed proteins between esophageal cancer cell lines and immortal cell line. Fifteen proteins were identified with differences of more than five folds, comprising the down-regulation of annexin A2, histone deacetylase 10 isoform beta and protein disulfide-isomerase ER-60 precursor, and the up-regulation of heat shock 70 kDa protein 9B precursor, solute carrier family 44 Member 3, heterogeneous nuclear ribonucleoprotein L (hnRNP L), eukaryotic translation initiation factor 4A isoform 2, triosephosphate isomerase1 (TPI), peroxiredoxin1 (PRX1), forminotransferase cyclodeaminase form (FTCD), fibrinogen gamma-A chain precursor, kinesin-like DNA binding protein, lamin A/C, cyclophilin A (CypA), and transcription factor MTSG1. Expression pattern of annexin A2 was verified by Western blotting, immunocytochemistry and immunohistochemistry analysis. The implication of these protein alterations correlated to the esophageal malignant transformation is discussed.
Assuntos
Carcinoma de Células Escamosas/química , Transformação Celular Neoplásica/química , Neoplasias Esofágicas/química , Proteínas de Neoplasias/análise , Proteômica , Anexinas/metabolismo , Western Blotting , Carcinoma de Células Escamosas/patologia , Adesão Celular , Linhagem Celular Tumoral , Eletroforese em Gel Bidimensional , Neoplasias Esofágicas/patologia , Humanos , Imuno-HistoquímicaRESUMO
The present study was designed to assess the synergistic antitumor effects of anthracenylmethyl homospermidine (ANTMHspd), a novel polyamine conjugate, with alpha-difluoromethylornithine (DFMO) and to elucidate the mechanism of these effects on human leukemia HL60 cells. Cell proliferation was assessed by the MTT assay. Cell cycle, apoptosis and mitochondria membrane potential (MMP) were evaluated by flow cytometry. Caspases and cytochrome c were detected by Western Blot analysis. The combination treatment strongly inhibited cell proliferation, induced cell apoptosis and caused an accumulation in the G1 phase with an accompaniment decrease in S phase. Moreover, reduction of MMP, release of cytochrome c and activation of caspase-3 and caspase-9 but not caspase-8 were observed during the combination-mediated apoptosis. All these findings demonstrated that the combination treatment with DFMO and ANTMHspd resulted in synergistic antitumor effects on HL60 cells.
Assuntos
Antracenos/farmacologia , Antineoplásicos/farmacologia , Eflornitina/farmacologia , Espermidina/análogos & derivados , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Proteínas Reguladoras de Apoptose/fisiologia , Western Blotting , Inibidores de Caspase , Caspases/metabolismo , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocromos c/metabolismo , Sinergismo Farmacológico , Inibidores Enzimáticos/farmacologia , Citometria de Fluxo , Fluoresceína-5-Isotiocianato , Corantes Fluorescentes , Células HL-60 , Humanos , Potenciais da Membrana/efeitos dos fármacos , Membranas Mitocondriais/efeitos dos fármacos , Espermidina/farmacologiaRESUMO
OBJECTIVE: To investigate the apoptosis-inducing effects of NNAMB, a novel polyamine conjugate, in erythroleukemia K562 cells and its molecular mechanism. METHODS: Cell viability was assessed by MTT assay and trypan blue dye exclusion method. The cell morphology was observed by fluorescence microscopy. The cell cycle distribution, apoptosis and mitochondrial membrane potential were measured by flow cytometry. The expression of caspase-3, -8, -9, cytochrome c in the K562 cells was detected by Western blot. RESULTS: NNAMB inhibited the proliferation of K562 cells. The cells treated with NNAMB showed a typical apoptotic morphology, Sub-G1 peak and loss of mitochondrial membrane potential. Western blot assay showed that NNAMB increased the expression of caspase-3, -9, cytochrome c but not caspase-8 in a dose-and time-dependent manner. CONCLUSION: NNAMB induces apoptosis via mitochondrial pathway in K562 cells.
Assuntos
Antracenos/farmacologia , Apoptose/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Poliaminas/farmacologia , Espermidina/análogos & derivados , Espermidina/farmacologia , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocromos c/metabolismo , Humanos , Células K562RESUMO
The title compound, [Ni(C(2)O(4))(C(10)H(8)N(2))](n), is isostructural with its Mn(II), Fe(II), Cu(II) and Zn(II) analogues. Each Ni(II) atom is chelated by two oxalate ligands and one 2,2'-bipyridine, forming a slightly distorted octa-hedral geometry. Oxlate acts as a bridge to link neighbouring pairs of Ni(II) cations, forming a one-dimensional wave-like chain. The crystal showed partial inversion twinning.
RESUMO
In the centrosymmetric title compound, [Gd(2)(C(7)H(3)F(2)O(2))(6)(C(10)H(8)N(2))(2)], the asymmetric unit comprises one cation chelated by two 2,5-difluoro-benzoate and one 2,2'-bipyridine. Two cations are linked into dimers via three bridging carboxyl-ate groups from three 2,5-difluoro-benzoic acid units. The Gd(III) ion is nine-coord-inated by seven O atoms and two N atoms.
RESUMO
The title compound, [Co(C(10)H(12)NO)(3)], was synthesized from cobalt(III) fluoride and 2-(propyl-imino-meth-yl)phenol in refluxing methanol. The Co(III) ion is hexa-coordinated by three N and three O atoms from three bidentate Schiff base ligands in an octa-hedral geometry.
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The pleiotropic pro-inflammatory cytokine, macrophage migration inhibitory factor (MIF), represents an important link between chronic inflammation and tumorigenesis. Although accumulating evidence demonstrates that MIF overexpression is implicated in the development and progression of multiple cancers, including esophageal squamous cell carcinoma (ESCC), the molecular mechanisms underlying its tumor-promoting roles in ESCC remain unclear. In the present study, we observed that MIF is overexpressed in ESCC and correlated significantly with lymph node metastasis, advanced clinical stage, and poor survival of ESCC. MIF knockdown attenuated the proliferation, migration, and invasion of ESCC cells in vitro and in vivo. Moreover, blockage of MIF expression decreased the activation of the Akt, MEK/ERK, and NF-κB pathways and enhanced sensitivity to apoptosis. Meanwhile, repression of MIF expression resulted in activation of glycogen synthase kinase 3 beta (GSK3ß) and subsequent decrease of active ß-catenin, as well as its downstream targets including cyclin D1, matrix metalloproteinase (MMP)-7, c-myc, and c-Jun. Collectively, our results provided mechanistic insights into the tumor-promoting role of MIF in ESCC, and suggested that MIF represents a potential therapeutic target for treatment of ESCC.
Assuntos
Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/patologia , Glicogênio Sintase Quinase 3 beta/fisiologia , Fatores Inibidores da Migração de Macrófagos/fisiologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Adulto , Idoso , Animais , Apoptose , Carcinoma de Células Escamosas/mortalidade , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal , Neoplasias Esofágicas/mortalidade , Carcinoma de Células Escamosas do Esôfago , Feminino , Humanos , Fatores Inibidores da Migração de Macrófagos/análise , Masculino , Camundongos , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de NeoplasiasRESUMO
Combining MS-based proteomic data with network and topological features of such network would identify more clinically relevant molecules and meaningfully expand the repertoire of proteins derived from MS analysis. The integrative topological indexes representing 95.96% information of seven individual topological measures of node proteins were calculated within a protein-protein interaction (PPI) network, built using 244 differentially expressed proteins (DEPs) identified by iTRAQ 2D-LC-MS/MS. Compared with DEPs, differentially expressed genes (DEGs) and comprehensive features (CFs), structurally dominant nodes (SDNs) based on integrative topological index distribution produced comparable classification performance in three different clinical settings using five independent gene expression data sets. The signature molecules of SDN-based classifier for distinction of early from late clinical TNM stages were enriched in biological traits of protein synthesis, intracellular localization and ribosome biogenesis, which suggests that ribosome biogenesis represents a promising therapeutic target for treating ESCC. In addition, ITGB1 expression selected exclusively by integrative topological measures correlated with clinical stages and prognosis, which was further validated with two independent cohorts of ESCC samples. Thus the integrative topological analysis of PPI networks proposed in this study provides an alternative approach to identify potential biomarkers and therapeutic targets from MS/MS data with functional insights in ESCC.
Assuntos
Biomarcadores Tumorais/biossíntese , Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , Proteínas de Membrana/biossíntese , Proteínas de Neoplasias/biossíntese , Mapas de Interação de Proteínas/genética , Proteínas Adaptadoras de Transdução de Sinal , Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Masculino , Proteínas de Membrana/genética , Terapia de Alvo Molecular , Proteínas de Neoplasias/genética , Estadiamento de Neoplasias , Prognóstico , Proteômica , Ribossomos/genética , Espectrometria de Massas em TandemRESUMO
BACKGROUND: In prokaryotic organisms, the mechanism responsible for the accurate partition of newly replicated chromosomes into daughter cells is incompletely understood. Segregation of the replication terminus of the circular prokaryotic chromosome poses special problems that have not previously been addressed. The aim of this study was to investigate the roles of several protein components (MreB, MreC, and MreD) of the prokaryotic cytoskeleton for the faithful transmission of the chromosomal terminus into daughter cells. METHODS: Strain LQ1 (mreB::cat), LQ2 (mreC::cat), and LQ3 (mreD::cat) were constructed using the Red recombination system. LQ11/pLAU53, LQ12/pLAU53, LQ13/pLAU53, LQ14/pLAU53, and LQ15/pLAU53 strains were generated by P1transduction of (tetO) 240 -Gm and (lacO) 240 -Km cassettes from strains IL2 and IL29. Fluorescence microscopy was performed to observe localization pattern of fluorescently-labeled origin and terminus foci in wild-type and mutant cells. SOS induction was monitored as gfp fluorescence from PsulA-gfp in log phase cells grown in Luria-Bertani medium at 37°C by measurement of emission at 525 nm with excitation at 470 nm in a microplate fluorescence reader. RESULTS: Mutational deletion of the mreB, mreC, or mreD genes was associated with selective loss of the terminus region in approximately 40% of the cells within growing cultures. This was accompanied by significant induction of the SOS DNA damage response, suggesting that deletion of terminus sequences may have occurred by chromosomal cleavage, presumably caused by ingrowth of the division septum prior to segregation of the replicated terminal. CONCLUSIONS: These results imply a role for the MreBCD cytoskeleton in the resolution of the final products of terminus replication and/or in the specific movement of newly replicated termini away from midcell prior to completion of septal ingrowth. This would identify a previously unrecognized stage in the overall process of chromosome segregation.
Assuntos
Citoesqueleto/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Segregação de Cromossomos/genética , Segregação de Cromossomos/fisiologiaRESUMO
OBJECTIVE: To investigate the correlation between sensitivity to tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and expression level of death receptor 5 (DR5) on tumor cell surface. METHODS: Anti-DR5 mAb was used to detect expression level of DR5 on surface of tumor cells by flow cytometry. Sensitivity to apoptosis induced by TRAIL was determined by TRAIL apoptosis kit. The correlation between expression level of DR5 and sensitivity to TRAIL was analyzed. RESULTS: The expression of DR5 on surface of tumor cells was approximately 97.9% in U937 cells, 95.1% in Jurkat cells, 93.8% in SW480 cells, 86.2% in HCT116 cells, 64.2% in HL-60 cells, 46.6% in HeLa cells and 13.1% in K562 cells, respectively. The apoptosis rate induced by TRAIL was 72.6% in U937 cells, 85.2% in Jurkat cells, 78.6% in SW480 cells, 70.2% in HCT116 cells, 60.1% in HL-60 cells, 45.4% in HeLa cells and 12.3% in K562 cells, respectively. There was a significant positive correlation between the expression level of DR5 with TRAIL-inducing apoptosis (r = 0.997, P < 0.001). CONCLUSION: TRAIL-inducing apoptosis is related to the expression level of DR5 on surface of tumor cells. The results confirm the importance of DR5 expression for induction of apoptosis by TRAIL.
Assuntos
Anticorpos Monoclonais/imunologia , Proteínas Reguladoras de Apoptose/farmacologia , Apoptose/efeitos dos fármacos , Glicoproteínas de Membrana/farmacologia , Receptores do Fator de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Humanos , Células Jurkat , Glicoproteínas de Membrana/metabolismo , Receptores do Ligante Indutor de Apoptose Relacionado a TNF , Receptores do Fator de Necrose Tumoral/imunologia , Ligante Indutor de Apoptose Relacionado a TNF , Fator de Necrose Tumoral alfa/metabolismo , Células U937RESUMO
OBJECTIVES: To evaluate the relationship between the functional integrity of sperm membrane and seminal parameters related to CASA. METHODS: Thirty-eight fertile and one hundrend and twenty four infertile males were tested the functional integrity of sperm membrane by the kit and parameters by CASA. RESULTS: There was a significant difference in the functional integrity of sperm membrane between fertile and infertile group (P < 0.01). The items related to CASA between normal and abnormal group in the functional integrity of sperm membrane had a remarkable difference, except motion degree, seminal volume and pH. CONCLUSIONS: To determine the functional integrity of sperm membrane can be used as a necessary supplementary method for CASA, and it has clinical significance in diagnosing, treating and researching male infertility.
Assuntos
Sêmen/fisiologia , Espermatozoides/fisiologia , Adulto , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Membranas/fisiologiaRESUMO
AIMS: The asymptomatic nature of early-stage esophageal squamous cell carcinoma (ESCC) results in late presentation and consequent dismal prognosis This study characterized 14-3-3σ protein expression in the multi-stage development of ESCC and determined its correlation with clinical features and prognosis. MATERIALS AND METHODS: Western blot was used to examine 14-3-3σ protein expression in normal esophageal epithelium (NEE), low grade intraepithelial neoplasia (LGIN), high grade intraepithelial neoplasia (HGIN), ESCC of TNM I to IV stage and various esophageal epithelial cell lines with different biological behavior. Immunohistochemistry was used to estimate 14-3-3σ protein in 110 biopsy samples of NEE, LGIN or HGIN and in 168 ESCC samples all of whom had follow-up data. Support vector machine (SVM) was used to develop a classifier for prognosis. RESULTS: 14-3-3σ decreased progressively from NEE to LGIN, to HGIN, and to ESCC. Chemoresistant sub-lines of EC9706/PTX and EC9706/CDDP showed high expression of 14-3-3σ protein compared with non-chemoresistant ESCC cell lines and immortalized NEC. Furthermore, the downregulation of 14-3-3σ correlated significantly with histological grade (Pâ=â0.000) and worse prognosis (Pâ=â0.004). Multivariate Cox regression analysis indicated that 14-3-3σ protein (Pâ=â0.016) and T stage (Pâ=â0.000) were independent prognostic factors for ESCC. The SVM ESCC classifier comprising sex, age, T stage, histological grade, lymph node metastasis, clinical stage and 14-3-3σ, distinguished significantly lower- and higher-risk ESCC patients (91.67% vs. 3.62%, Pâ=â0.000). CONCLUSIONS: Downregulation of 14-3-3σ arises early in the development of ESCC and predicts poor survival, suggesting that 14-3-3σ may be a biomarker for early detection of high-risk subjects and diagnosis of ESCC. Our seven-feature SVM classifier for ESCC prognosis may help to inform clinical decisions and tailor individual therapy.
Assuntos
Proteínas 14-3-3/metabolismo , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patologia , Idoso , Western Blotting , Carcinoma de Células Escamosas do Esôfago , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Máquina de Vetores de SuporteRESUMO
This study was purposed to explore the effect of hyperactivation of c-Jun NH(2)-terminal protein kinase (JNK) on the proliferation of B lymphoma cells. The human B lymphoma cell lines Daudi and Raji were chosen as research objects. The expression of JNK protein was determined by Western blot. The subcellular localization of JNK protein was detected by immunofluorescence. The cell cycle was analyzed by flow cytometry. The suppressive effect of JNK inhibitor SP600125 on the proliferation of Daudi and Raji cells was assayed by ATPLite method. The results demonstrated that hyperactivation of JNK has been found in Daudi and Raji cells. Immunofluorescence confirmed the aberrant subcellular localization of JNK protein in Daudi and Raji cells. Cell cycle assay revealed that Daudi and Raji cells underwent G(2)-M arrest in the presence of SP600125. Furthermore, Daudi and Raji cells showed significant increase in sub-G(1) population, an indicator of apoptotic cells, with the treatment of JNK inhibitors. These data suggested that JNK inhibitors suppressed the growth of B lymphoma cells via cell cycle arrest and apoptosis. Daudi and Raji cells treated with different concentrations of JNK selective inhibitor SP600125 showed dose-dependent reduction in the growth of Daudi and Raji cells. It is concluded that hyperactivation of JNK enhance the proliferation of Daudi and Raji cells. The aberrant subcellular localization of JNK protein may facilitate the nuclear accumulation of basal JNK activity, which made JNK to be a potential target to treat human B lymphoma.
Assuntos
Proliferação de Células , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Linfoma de Células B/patologia , Linhagem Celular Tumoral , Humanos , Linfoma de Células B/metabolismoRESUMO
The striking 3-4:1 male predominance of esophageal squamous cell carcinoma (ESCC) has not yet been well explained. Our hypothesis is that the changes in level of estrogen and/or subtype of estrogen receptor (ER) may exert a protective factor in esophageal carcinogenesis and prognosis of ESCC. Radioimmunoassay (RIA) was used to determine the serum level of estradiol in healthy cohort from high-incidence area (HIA) and low-incidence area (LIA) for esophageal cancer as well as patients with ESCC from HIA in Henan, northern China. The ERß expression profiling during the multi-stage progression of ESCC pathogenesis was evaluated by immunohistochemistry (IHC). Both males and females from HIA had significant decreases of serum estradiol in high-risk subjects predisposing for ESCC compared to healthy counterparts from LIA (P < 0.01). Furthermore, patients with ESCC from HIA developed the lowest level of estradiol (P < 0.01). ERß expressed in precursor lesions of ESCC and changed quantitatively and qualitatively with disease progression during the multi-stages process of esophageal carcinogenesis. High frequency of ERß expression was correlated with less aggressive potential of clinical behavior (P = 0.012, 0.015 for lymph node metastasis and tumor stage, respectively). This study indicates that lower serum level of estradiol may represent higher predisposition for development of ESCC, and ERß expression and/or nuclear location may predict better outcome for patients with ESCC. The present results provide clues to explain the striking gender difference for ESCC, which warrants further investigations on potential applications of estrogen or analogs in prevention of ESCC.
Assuntos
Carcinoma de Células Escamosas/sangue , Neoplasias Esofágicas/sangue , Estradiol/sangue , Receptor beta de Estrogênio/sangue , Adulto , Carcinoma de Células Escamosas/diagnóstico , Estudos de Casos e Controles , China , Neoplasias Esofágicas/diagnóstico , Esôfago/metabolismo , Feminino , Humanos , Técnicas Imunoenzimáticas , Incidência , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , PrognósticoRESUMO
In the murine splenocytes, CD8α+ dendritic cells (abbreviated as 8+DC) and CD8α- dendritic cells (abbreviated as 8-DC) are identified with some vague features for each of them. 8+DCs but not 8-DCs cross-prime cytotoxic T cells in vivo. We aim to distinguish the two subtypes of DC based on gene expression profiling. Suppressive subtractive hybridization was undertaken to get differentially expressed genes from such subtracted cDNA library specific to 8+DC. A total of 114 sequences from the subtracted cDNA library specific to 8+DC library were analyzed. Most of them are known proteins, but some of them were novel, either totally novel genes or homologs to known genes, but with novel exon. About 55 probably novel exons were discovered, and 11 exons had longer length than those in gene bank. The clones 12, 44, 79, and 110 have no match with known sequences in gene bank. Then, semi-quantitative PCR was done to compare the expression of the enriched sequences between 8+DC and 8-DC. About 14 genes are differentially expressed in 8+DC. Therefore, SSH is an effective method to clone differentially expressed genes for 8+DC compared to 8-DC.
Assuntos
Antígenos CD8 , Células Dendríticas/imunologia , Regulação da Expressão Gênica/imunologia , Baço/imunologia , Animais , Células Dendríticas/citologia , Feminino , Perfilação da Expressão Gênica , Masculino , Camundongos , Baço/citologiaRESUMO
An agonistic antibody against TNF-related apoptosis-inducing ligand death receptor 5 (DR5) is a practicable candidate drug for antitumor therapy. In this study, a novel murine anti-human DR5 monoclonal antibody, mDRA-6(IgG1-κ), has been generated. This study aimed to explore the caspase-dependent and mitochondrial mechanisms of mDRA-6 in inducing apoptosis in human leukemia Jurkat cells. The apoptotic effects of mDRA-6 on Jurkat cells, which express DR5 on the cell surface, were detected by flow cytometry and western blot after exposure to different doses of mDRA-6 and at fixed doses of mDRA-6 at different times. It was demonstrated that mDRA-6 can induce Jurkat cell apoptosis via caspase- and mitochondrial-dependent pathways. These results indicate that the novel antibody mDRA-6 against DR5 has an antitumor function and may provide a new reagent for tumor therapy.