RESUMEN
BACKGROUND: Cryopreservation process negatively affects spermatozoa functions. Humanin, a small polypeptide encoded in the mitochondrial genome, is well known for its role in cell survival. OBJECTIVE: To quantify the endogenous levels of humanin in seminal plasma of crossbred Frieswal bulls and to study its role in cryoprotection. The presence of humanin in bull spermatozoa was also investigated. MATERIALS AND METHODS: A total of 40 semen samples were separated into two groups based on the initial progressive motility (IPM): Good (IPM >70%) and Poor (IPM <50%) groups; and/or based on the post-thaw motility (PTM): Freezable (PTM>50%) and Non-freezable (PTM < 50%) groups. Humanin concentration in seminal plasma (SP-HN) was quantified using ELISA. RESULTS: SP-HN concentration ranged from undetectable to 67.6 pg/mL with a median level of 35.2 pg/mL. SP-HN level was significantly higher in the good quality semen group than in the poor quality semen group (p<0.001), and also significantly higher in the freezable group than in the non-freezable group (p<0.001). SP-HN level was positively correlated with initial progressive motility, post-thaw semen motility, viability, acrosome intactness and plasma membrane integrity, but negatively correlated the level of reactive oxygen species and malondialdehyde content. Immunochemical localization showed the presence of humanin in the proximal region of the middle piece of spermatozoa. CONCLUSION: Endogenous humanin level had significant correlation with semen quality and might protect sperm cells against freeze-induced oxidative stress. doi.org/10.54680/fr22510110712.
Asunto(s)
Preservación de Semen , Semen , Masculino , Animales , Bovinos , Análisis de Semen , Criopreservación/veterinaria , Preservación de Semen/veterinaria , Espermatozoides/química , Péptidos y Proteínas de Señalización Intracelular/análisis , Motilidad EspermáticaRESUMEN
BACKGROUND: Humanin is an endogenous mitochondria-derived peptide that plays critical roles in oxidative stress, inflammation and CAD. In this study, we measured the levels of circulating humanin, markers of oxidative stress and inflammation in patients with unstable angina and MI and studied the relationship between these parameters and major adverse cardiac events (MACE). METHODS: A total of 327 subjects were recruited from the inpatient department at First Hospital of Jilin University and divided into 3 groups [control, angina and myocardial infarction (MI)] based on the clinical data and the results of the angiography. Serum humanin and thiobarbituric acid reactive substances (TBARS) were measured at the time of initial admission. The hospitalization data and MACE of all patients were collected. RESULTS: Circulating humanin levels were lower in the angina group compared to controls [124.22⯱â¯63.02 vs. 157.77⯱â¯99.93â¯pg/ml, pâ¯<â¯0.05] and even lower in MI patients [67.17⯱â¯24.35â¯pg/ml, pâ¯<â¯0.05 vs controls] and oxidative stress marker were higher in MI patients compared to the control and angina groups [12.94⯱â¯4.55 vs. 8.26⯱â¯1.66 vs. 9.06⯱â¯2.47 umol/ml, pâ¯<â¯0.05]. Lower circulating humanin levels was an independent risk factor of MI patients. Circulating humanin levels could be used to predict MACE in angina group. CONCLUSIONS: Lower circulating humanin levels was an independent risk factor for CAD, and a potential prognostic marker for mild CAD. GENERAL SIGNIFICANCE: Humanin may become a new index for the diagnosis and treatment of CAD.
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Enfermedad de la Arteria Coronaria/metabolismo , Péptidos y Proteínas de Señalización Intracelular/análisis , Adulto , Angina de Pecho/metabolismo , Angina de Pecho/fisiopatología , Biomarcadores/sangre , Enfermedad de la Arteria Coronaria/sangre , Femenino , Corazón , Humanos , Inflamación/metabolismo , Péptidos y Proteínas de Señalización Intracelular/sangre , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Persona de Mediana Edad , Mitocondrias , Infarto del Miocardio/metabolismo , Estrés Oxidativo/fisiología , Pronóstico , Factores de RiesgoRESUMEN
Background: Tumor necrosis factor (TNF)-alpha-induced protein 8-like 2 (TIPE2 or TNFAIP8L2) is a newly discovered negative immune regulator. Studies have shown that TIPE2 causes significant malignant biological effects and is differentially expressed in various malignant tumors. However, the expression and roles of TIPE2 in pancreatic ductal adenocarcinoma (PDAC) are largely unknown. Materials and Methods: The expression of TIPE2 in PDAC tissues was assessed by immunohistochemistry, qPCR and western blot analysis and related clinicopathological parameters including survival time were analyzed. After overexpression of TIPE2, cell proliferation and apoptosis analysis were conducted, and the associated underlying molecular mechanism was also explored. Results: In the present study, TIPE2 was upregulated in early PDAC tissues, and TIPE2 expression decreased as the tumor progressed (P<0.001). TIPE2 expression was negatively associated with tumor size, TNM stage and metastasis of lymph nodes. Furthermore, as an independent risk factor, TIPE2 could be used to predict the survival of patients with PDAC (P=0.035). TIPE2 overexpression significantly suppressed the viability, proliferation and induced apoptosis of PDAC cells by inhibiting survivin and increasing the activity of caspase3/7. Conclusions: For the first time, this study demonstrated that TIPE2 is an independent prognostic factor in PDAC. TIPE2 inhibited the proliferation and induced apoptosis via regulating survivin/caspase3/7 signaling pathway. These results indicated that TIPE2 is a potential biomarker for predicting the prognosis of PDAC patients and plays a pivotal role in the progression of PDAC.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Carcinoma Ductal Pancreático/mortalidad , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Neoplasias Pancreáticas/mortalidad , Apoptosis/genética , Biomarcadores de Tumor/análisis , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/cirugía , Caspasa 3/metabolismo , Línea Celular Tumoral , Proliferación Celular/genética , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Regulación Neoplásica de la Expresión Génica , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Invasividad Neoplásica/genética , Páncreas/patología , Páncreas/cirugía , Pancreatectomía , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/cirugía , Pronóstico , Transducción de Señal/genética , Survivin/metabolismo , Regulación hacia ArribaRESUMEN
Senescence marker protein 30 (SMP30) is a cell survival factor playing an important role in vitamin C synthesis and antiapoptosis. Moreover, its cytoprotective role suggests a possibility to be related to cancer cell survival. Mammary carcinoma is a common cancer in both humans and animals. Because of its histopathological diversity, especially in the early stage, histopathological diagnosis may be complicated; therefore, a diagnostic marker is helpful for confirmation. The present study analyzed the expression pattern of SMP30 in mammary carcinoma in humans, dogs, and cats. Immunohistochemistry, immunofluorescence, and western blot analysis were used to investigate SMP30 expression patterns. The expression was specifically observed in neoplastic glandular epithelial cells. The expression increased with the malignancy of glandular epithelial cells with a highly proliferative status. However, SMP30 expression was low in normal mammary gland tissues or well-differentiated adenoma tissues. The patterns were consistently reproduced in canine primary mammary carcinoma cells and MCF-7 and MDA-MB-231 human carcinoma cell lines. This study provides useful information to understand SMP30 expression in various stages of mammary carcinoma and to suggest its utility as a pan-species diagnostic marker, thereby helping to establish strategies for diagnosing mammary carcinoma in several species.
Asunto(s)
Neoplasias de la Mama/patología , Proteínas de Unión al Calcio/análisis , Enfermedades de los Gatos/patología , Enfermedades de los Perros/patología , Péptidos y Proteínas de Señalización Intracelular/análisis , Neoplasias Mamarias Animales/patología , Animales , Biomarcadores de Tumor/análisis , Mama/patología , Neoplasias de la Mama/diagnóstico , Enfermedades de los Gatos/diagnóstico , Gatos , Línea Celular Tumoral , Enfermedades de los Perros/diagnóstico , Perros , Femenino , Humanos , Células MCF-7 , Neoplasias Mamarias Animales/diagnóstico , PronósticoRESUMEN
AIMS: To investigate novel biomarker for diagnosis of cervical cancer, we analyzed the datasets in Gene Expression Omnibus (GEO) and confirmed the candidate biomarker in patient sample. MATERIALS AND METHODS: We collected major datasets of cervical cancer in GEO, and analyzed the differential expression of normal and cancer samples online with GEO2R and tested the differences, then focus on the GSE63514 to screen the target genes in different histological grades by using the R-Bioconductor package and R-heatmap. Then human specimens from the cervix in different histological grades were used to confirm the top 8 genes expression by immunohistochemical staining using Ki67 as a standard control. RESULTS: We identified genes differentially expressed in normal and cervical cancer, 274 upregulated genes and 206 downregulated genes. After intersection with GSE63514, we found the obvious tendency in different histological grades. Then we screened the top 24 genes, and confirmed the top 8 genes in human cervix tissues. Immunohistochemical (IHC) results confirmed that keratin 17 (KRT17) was not expressed in normal cervical tissues and was over-expressed in cervical cancer. Cysteine-rich secretory protein-2 (CRISP2) was less expressed in high-grade squamous intraepithelial lesions (HSILs) than in other histological grades. CONCLUSION: For the good repeatability and consistency of KRT17 and CRISP2, they may be good candidate biomarkers. Combined analysis of KRT17, CRISP2 expression at both genetic and protein levels can determine different histological grades of cervical squamous cell carcinoma. Such combined analysis is capable of improving diagnostic accuracy of cervical cancer.
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Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/genética , Moléculas de Adhesión Celular/genética , Queratina-17/genética , Displasia del Cuello del Útero/genética , Neoplasias del Cuello Uterino/genética , Biomarcadores de Tumor/análisis , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/patología , Moléculas de Adhesión Celular/análisis , Proteínas de Ciclo Celular/análisis , Proteínas de Ciclo Celular/genética , Cuello del Útero/metabolismo , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/genética , Bases de Datos Genéticas , Conjuntos de Datos como Asunto , Desmogleína 1/análisis , Desmogleína 1/genética , Regulación hacia Abajo , Femenino , Perfilación de la Expresión Génica/métodos , Marcadores Genéticos , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/genética , Queratina-17/análisis , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Clasificación del Tumor , Proteínas de Neurofilamentos/análisis , Proteínas de Neurofilamentos/genética , Proteínas y Péptidos Salivales/análisis , Proteínas y Péptidos Salivales/genética , Proteínas de Plasma Seminal/análisis , Proteínas de Plasma Seminal/genética , Regulación hacia Arriba , Neoplasias del Cuello Uterino/química , Neoplasias del Cuello Uterino/patología , Displasia del Cuello del Útero/química , Displasia del Cuello del Útero/patologíaRESUMEN
The widespread use of silver nanoparticles (AgNPs), their many sources for human exposure, and the ability of AgNPs to enter organisms and induce general toxicological responses have raised concerns regarding their public health and environmental safety. To elucidate the differential toxic effects of polyvinylpyrrolidone-capped AgNPs with different primary particle sizes (i.e. 5, 50, and 75 nm), we performed a battery of cytotoxicity and genotoxicity assays and examined the inflammatory responses in two human cell lines (i.e. HepG2 and A549). Concentration-dependent decreases in cell proliferation and mitochondrial membrane potential and increases in cytokine (i.e. interleukin-6 and interleukin-8) excretion indicated disruption of mitochondrial function and inflammation as the main mediating factors of AgNPs-induced cytotoxicity. An incremental increase in genotoxicity with decreasing AgNPs diameter was noted in HepG2 cells, which was associated with S and G2/M accumulation and transcriptional activation of the GADD45α promoter as reflected by luciferase activity. Dose-related genetic damage, as indicated by Olive tail moment and micronucleus formation, was also observed in A549 cells, but these effects as well as the AgNPs-induced cytotoxicity were more associated with ionic Ag release from nanoparticles (NPs). In summary, the present study addressed different toxicity mechanisms of AgNPs, depending on the cell model, toxicological endpoint, particle size, and degree of Ag+ release from NPs. The results suggest that the GADD45α promoter-driven luciferase reporter cell system provided a rapid screening tool for the identification of genotoxic properties of NPs across a range of different sizes and concentrations.
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Nanopartículas del Metal/efectos adversos , Mutágenos/análisis , Povidona/efectos adversos , Plata/efectos adversos , Células A549 , Línea Celular , Ensayo Cometa , Citotoxinas/análisis , Relación Dosis-Respuesta a Droga , Células Hep G2 , Humanos , Inflamación , Péptidos y Proteínas de Señalización Intracelular/análisis , Luciferasas/análisis , Tamaño de la Partícula , Proteinas GADD45RESUMEN
We previously reported that a strong immunoreactivity of tripartite motif-containing 44 (TRIM44) predicts the poor prognosis of patients with invasive breast cancer, and proposed that TRIM44 activates nuclear factor-κB (NF-κB) signaling as a causative mechanism. In the present study, we examined the clinicopathological roles of A20, which is known to be an NF-κB responsive gene, with TRIM44, in an updated cohort. Tissue samples of invasive breast cancer were obtained from 140 Japanese female breast cancer patients who underwent surgical treatment. Immunoreactivities of A20 and TRIM44 were analyzed using specific antibodies for each protein. A positive A20 immunoreactivity was significantly associated with a shorter disease-free survival (P = 0.043) and was positively correlated with TRIM44 immunoreactivity (P = 0.039). Combined use of the immunoreactivities for two proteins revealed that double-positive status for both A20 and TRIM44 immunoreactivities was associated with a shorter disease-free survival (P = 0.012) and was an independent factor for poor prognosis. These results indicate that a combined A20 and TRIM44 immunoreactivity predicted the prognosis of patients with invasive breast cancer. Moreover, the positive correlation between A20 and TRIM44 immunoreactivities suggested that the activation of NF-κB signaling by TRIM44 could occur in clinical breast cancer tissues.
Asunto(s)
Neoplasias de la Mama , Péptidos y Proteínas de Señalización Intracelular , Pronóstico , Proteínas de Motivos Tripartitos , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Femenino , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular/análisis , Japón , Proteínas de Motivos Tripartitos/análisis , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa/análisisRESUMEN
BACKGROUND: Pancreatic neuroendocrine tumors are rare neoplasms for which few predictive and/or prognostic biomarkers have been validated. Our previous work suggested the potential of the combined expression of N-myc downstream-regulated gen-1 (NDRG-1), O6-methylguanine DNA methyltransferase (MGMT) and Pleckstrin homology-like domain family A member 3 (PHLDA-3) as prognostic factors for relapse and survival. METHODS: In this new multicenter study we evaluated immunohistochemistry expression in 76 patients with advanced PanNET who were treated with capecitabine-temozolomide or everolimus. Based on the immunohistochemistry panel, an immunohistochemistry prognostic score (IPS) was developed. RESULTS: In patients treated with capecitabine and temozolomide, low IPS was an independent prognostic factor for progression-free-survival and overall-survival. Similar findings were observed with highest IPS for overall-survival in patients treated with everolimus. CONCLUSION: From our knowledge, it is the first time that a simple IPS could be useful to predict outcome for patients with metastatic pancreatic neuroendocrine tumors treated with everolimus or capecitabine and temozolomide.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Everolimus/uso terapéutico , Inmunohistoquímica/métodos , Inmunosupresores/uso terapéutico , Tumores Neuroendocrinos/tratamiento farmacológico , Tumores Neuroendocrinos/patología , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/patología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/análisis , Proteínas de Ciclo Celular/análisis , Línea Celular Tumoral , Metilasas de Modificación del ADN/análisis , Enzimas Reparadoras del ADN/análisis , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Tumores Neuroendocrinos/mortalidad , Proteínas Nucleares/análisis , Neoplasias Pancreáticas/mortalidad , Pronóstico , Supervivencia sin Progresión , Análisis de Supervivencia , Proteínas Supresoras de Tumor/análisis , Adulto JovenRESUMEN
BACKGROUND: Circular RNAs (circRNAs) act as vital regulators of gene expression in a variety of cancers. However, the role of circRNAs in gastric cancer (GC) remains largely unexplored. Herein, we identified that circTMEM87A sponges miR-142-5p to promote GC progression through up-regulating ULK1 expression. METHODS: The expression of circTMEM87A in GC was determined by RNA sequencing and quantitative real-time PCR (qRT-PCR). The effects of knockdown or exogenous expression of circTMEM87A on GC cell phenotypes were evaluated both in vitro and in vivo. The interacting miRNA of circTMEM87A was predicted by bioinformatics and confirmed by RNA pull-down, dual-luciferase reporter assay and fluorescence in situ hybridization (FISH). The mechanism by which circTMEM87A/miR-142-5p/ULK1 axis promotes GC was determined by western blot, GFP/mRFP-LC3 puncta analysis, transmission electron microscope (TEM). RESULTS: CircTMEM87A was dramatically elevated in GC tissues and cell lines, and high circTMEM87A expression was closely correlated with poor prognosis of GC patients. Knockdown of circTMEM87A suppressed cell growth, migration, invasion and induced apoptosis in vitro, as well as inhibited GC tumorigenicity and lung metastasis potential in vivo. Meanwhile, circTMEM87A overexpression had the opposite effects. Furthermore, we demonstrated that circTMEM87A could act as a sponge of miR-142-5p to regulate ULK1 expression and GC progression. CONCLUSIONS: Our findings suggest that circTMEM87A functions as an oncogene through the miR-142-5p/ULK1 axis in GC. CircTMEM87A might be a prognostic biomarker as well as a promising therapeutic target for GC.
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Homólogo de la Proteína 1 Relacionada con la Autofagia/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Péptidos y Proteínas de Señalización Intracelular/efectos de los fármacos , MicroARNs/efectos de los fármacos , ARN Circular/farmacología , Neoplasias Gástricas/etiología , Homólogo de la Proteína 1 Relacionada con la Autofagia/análisis , Homólogo de la Proteína 1 Relacionada con la Autofagia/genética , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Línea Celular Tumoral/efectos de los fármacos , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/genética , MicroARNs/análisis , MicroARNs/genética , ARN Circular/uso terapéutico , Neoplasias Gástricas/fisiopatologíaRESUMEN
OBJECTIVE: To investigate the relationships between diabetic nephropathy (DN) and insulin resistance, inflammation, thioredoxin (Trx), thioredoxin-interacting protein (Txnip), Cystatin C (CysC) and serum complement levels. PATIENTS AND METHODS: A total of 119 patients with type 2 diabetes mellitus (T2DM) treated in the Endocrinology Department of our hospital from January 2017 to December 2017 were enrolled as the experiment group, while 30 healthy volunteers were selected as the control group. The expression levels of inflammatory factors, Trx, Txnip, CysC and serum complements in every subject were detected. In addition, the type 2 diabetic nephropathy rat model was established via high-fat diet and injection of low-dose streptozotocin. Blood glucose, insulin resistance indexes and 24h-urinary albumin excretion were measured, and the histomorphological characteristics of the kidney in animals were observed. RESULTS: In clinical subjects, Trx level was notably lower in the simple DM group, early DN group and clinical DN group in comparison with that in the control group. The levels of Txnip and CysC in the simple DM group, early DN group and clinical DN group were remarkably higher than those in the control group. Moreover, the expression levels of TNF-α and IL-6 in the clinical DN group were significantly elevated compared with those in the simple DM group and early DN group. In addition, C1q expression in the clinical DN group was higher than that in the simple DM group and early DN group. In model rats, HOMA-IR was distinctly higher in the DM group and DN group than that in the control group. The ratio of kidney weight to body weight (KW/BW) was evidently higher in the DN group in comparison with that in the control group and DM group. CONCLUSIONS: Insulin resistance, inflammatory factors, and levels of Trx, Txnip, CysC and serum complement C1q are related to the progression of DM.
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Diabetes Mellitus Experimental/patología , Diabetes Mellitus Tipo 2/patología , Nefropatías Diabéticas/patología , Inflamación/patología , Animales , Glucemia/análisis , Proteínas Portadoras/análisis , Proteínas de Ciclo Celular/análisis , Complemento C1q/análisis , Cistatina C/análisis , Diabetes Mellitus Experimental/inducido químicamente , Diabetes Mellitus Tipo 2/inducido químicamente , Nefropatías Diabéticas/inducido químicamente , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Humanos , Inflamación/inducido químicamente , Resistencia a la Insulina , Péptidos y Proteínas de Señalización Intracelular/análisis , Masculino , Proteínas de la Membrana/análisis , Ratas , Ratas Sprague-Dawley , Estreptozocina/administración & dosificación , Tiorredoxinas/análisisRESUMEN
Niemann-Pick disease type C is a rare, fatal neurodegenerative disorder characterized by massive intracellular accumulation of cholesterol. In most cases, loss-of-function mutations in the NPC1 gene that encodes lysosomal cholesterol transporter NPC1 are responsible for the disease, and more than half of the mutations are considered to interfere with the biogenesis or folding of the protein. We previously identified a series of oxysterol derivatives and phenanthridine-6-one derivatives as pharmacological chaperones, i.e., small molecules that can rescue folding-defective phenotypes of mutated NPC1, opening up an avenue to develop chaperone therapy for Niemann-Pick disease type C. Here, we present an improved image-based screen for NPC1 chaperones and we describe its application for drug-repurposing screening. We identified some azole antifungals, including itraconazole and posaconazole, and a kinase inhibitor, lapatinib, as probable pharmacological chaperones. A photo-crosslinking study confirmed direct binding of itraconazole to a representative folding-defective mutant protein, NPC1-I1061T. Competitive photo-crosslinking experiments suggested that oxysterol-based chaperones and itraconazole share the same or adjacent binding site(s), and the sensitivity of the crosslinking to P691S mutation in the sterol-sensing domain supports the hypothesis that their binding sites are located near this domain. Although the azoles were less effective in reducing cholesterol accumulation than the oxysterol-derived chaperones or an HDAC inhibitor, LBH-589, our findings should offer new starting points for medicinal chemistry efforts to develop better pharmacological chaperones for NPC1.
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Descubrimiento de Drogas/métodos , Péptidos y Proteínas de Señalización Intracelular/genética , Enfermedad de Niemann-Pick Tipo C/tratamiento farmacológico , Pliegue de Proteína/efectos de los fármacos , Reposicionamiento de Medicamentos/métodos , Células HEK293 , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Mutación/efectos de los fármacos , Proteína Niemann-Pick C1 , Enfermedad de Niemann-Pick Tipo C/genética , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/farmacologíaRESUMEN
Aim: Novel biomarkers that are able to accurately monitor tuberculosis (TB) treatment effectiveness are needed to adjust therapy and identify a need for a regimen change. Materials & methods: In our study, conducted on a cohort comprising 100 pulmonary TB patients, we analyzed the role of plasma cytokines and Toll-like receptors expression as biomarkers of treatment response. Results: Changes in toll-interacting protein (TOLLIP) and lymphocyte antigen 96 (LY96) gene expression as well as nine cytokine levels over the first 2 months were significantly associated with successful treatment outcome. Successful treatment was associated with higher serum concentration of Toll-like receptor-2. Conclusion: Our results suggest that differential expression of specific effector molecules and dynamics of selected cytokines may help to identify those responding to TB treatment early.
Asunto(s)
Tuberculosis Pulmonar/tratamiento farmacológico , Adulto , Antituberculosos/uso terapéutico , Biomarcadores Farmacológicos/sangre , Estudios de Cohortes , Citocinas/sangre , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/sangre , Estudios Longitudinales , Antígeno 96 de los Linfocitos/análisis , Antígeno 96 de los Linfocitos/sangre , Masculino , Persona de Mediana Edad , Mycobacterium tuberculosis/patogenicidad , Resultado del Tratamiento , Tuberculosis/tratamiento farmacológico , Tuberculosis Pulmonar/sangre , Tuberculosis Pulmonar/inmunologíaRESUMEN
OBJECTIVES: The interferon-induced protein with multiple tetratricopeptide repeats 3 (IFIT3) seems to be associated with the prognosis in pancreatic cancer. Here we clarify whether the heterogeneity of IFIT3 expression affects previous IFIT3 analysis. METHODS: This retrospective study analyzes pancreatic cancer tissue samples retrieved by surgery from 2 independent patient cohorts. Patients underwent either primary surgery (n = 72) or received neoadjuvant chemotherapy (n = 12). Immunohistochemistry assessed IFIT3 expression and its heterogeneity. Complementarily, we analyzed publicly available transcriptomic data (n = 903). RESULTS: Of the primarily resected tumors, 16.4% were heterogeneous. Patients with IFIT3-negative tumors did not survive longer compared with patients with IFIT3-positive tumors. An analysis of publicly available data confirmed this result. Patients developing lung metastases had the best prognosis (4.8 years) with significantly lower IFIT3 expression compared with liver metastasis (P = 0.0117). Patients receiving neoadjuvant therapy who are IFIT3 negative had a longer disease-free survival (1.2 vs 0.3 years, P = 0.0081). CONCLUSIONS: Low IFIT3 expression is not associated with longer survival. Divergent results from tissue microarray analyses could be explained with tumor heterogeneity. As a single biomarker, IFIT3 is not suitable for predicting disease prognosis. Recurrence of lung metastases and response to neoadjuvant chemotherapy are associated with low IFIT3 expression.
Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma Ductal Pancreático/química , Péptidos y Proteínas de Señalización Intracelular/análisis , Neoplasias Pulmonares/química , Neoplasias Pancreáticas/química , Anciano , Biomarcadores de Tumor/genética , Carcinoma Ductal Pancreático/mortalidad , Carcinoma Ductal Pancreático/secundario , Carcinoma Ductal Pancreático/terapia , Quimioterapia Adyuvante , Bases de Datos Genéticas , Supervivencia sin Enfermedad , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Neoplasias Pulmonares/mortalidad , Neoplasias Pulmonares/secundario , Neoplasias Pulmonares/terapia , Masculino , Terapia Neoadyuvante , Recurrencia Local de Neoplasia , Pancreatectomía , Neoplasias Pancreáticas/mortalidad , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/terapia , Estudios Retrospectivos , Factores de TiempoRESUMEN
We screened cell line and plasma-derived exosomes for molecules that localize to mitochondria or that reflect mitochondrial integrity. SH-SY5Y cell-derived exosomes contained humanin, citrate synthase, and fibroblast growth factor 21 protein, and plasma-derived exosomes contained humanin, voltage-dependent anion-selective channel 1, and transcription factor A protein. Nuclear mitochondrial (NUMT) DNA complicated analyses of mitochondrial DNA (mtDNA), which otherwise suggested exosomes contain at most very low amounts of extended mtDNA sequences but likely contain degraded pieces of mtDNA. Cell and plasma-derived exosomes contained several mtDNA-derived mRNA sequences, including those for ND2, CO2, and humanin. These results can guide exosome-focused, mitochondria-pertinent biomarker development.
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Sangre/metabolismo , ADN Mitocondrial/análisis , Exosomas/química , Mitocondrias/química , Línea Celular , Cromatografía en Gel , Complejo IV de Transporte de Electrones/análisis , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Microscopía Electrónica de Transmisión , NADH Deshidrogenasa/análisisRESUMEN
Triple-negative breast cancer (TNBC) that lacks expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) is a breast cancer subtype with very aggressive metastasis and poor prognosis. Unique cartilage matrix-associated protein (UCMA) is a vitamin K-dependent protein (VKDP) with a high-density γ-carboxyglutamic acid (Gla) domain due to the action of vitamin K. UCMA promotes osteoblast differentiation and mineral deposition in bone and suppresses calcification in vessels. However, correlation between UCMA and TNBC is unknown. This study investigated the inhibitory effect of UCMA on TNBC cell in vitro migration, invasion, and colony formation in addition to in vivo tumorigenesis. Cell migration and invasion significantly decreased in Ucma-overexpressing MDA-MB-231 and 4T1 cells compared to the mock control cells. Also, colony formation and the number of colonies significantly decreased in Ucma-overexpressing MDA-MB-231 and 4T1 cells. These results indicate that UCMA significantly inhibits the migration, invasion, and colony formation of TNBC cells. In an in vivo xenograft mouse model, tumor growth significantly decreased in mice bearing Ucma-overexpressing TNBC cells compared to the mock control cells, indicating that UCMA reduced in vivo tumor growth, similar to the inhibitory role of UCMA in vitro. Survival analysis using publicly available database showed that high UCMA expression significantly correlated with favorable relapse-free survival in TNBC patients compared to those with the other VKDPs, matrix Gla protein (MGP) and osteocalcin (OCN). Collectively, this study suggests that UCMA is a promising new therapeutic agent for TNBC.
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Péptidos y Proteínas de Señalización Intracelular/metabolismo , Invasividad Neoplásica/patología , Neoplasias de la Mama Triple Negativas/patología , Animales , Línea Celular Tumoral , Movimiento Celular , Femenino , Humanos , Péptidos y Proteínas de Señalización Intercelular , Péptidos y Proteínas de Señalización Intracelular/análisis , Ratones Endogámicos BALB C , Neoplasias de la Mama Triple Negativas/metabolismoRESUMEN
Inhibition of glycolysis has been considered as a therapeutic approach in aggressive cancers including lung cancer. Abbreviated gluconeogenesis, mediated by phosphoenolpyruvate carboxykinase (PEPCK), was recently discovered to partially circumvent the need for glycolysis in lung cancer cells. However, the interplay of glycolysis and gluconeogenesis in lung cancer is still poorly understood. Here, we analyzed the expression of GLUT1, the prime glucose transporter, and of PCK1 and PCK2, the cytoplasmic and mitochondrial isoforms of PEPCK, in 450 samples of non-small cell lung cancer (NSCLC) and in 54 NSCLC metastases using tissue microarrays and whole tumor sections. Spatial distribution was assessed by automated image analysis. Additionally, glycolytic and gluconeogenic gene expression was inferred from The Cancer Genome Atlas (TCGA) datasets. We found that PCK2 was preferentially expressed in the lung adenocarcinoma subtype, while GLUT1 expression was higher in squamous cell carcinoma. GLUT1 and PCK2 were inversely correlated, GLUT1 showing elevated expression in larger tumors while PCK2 was highest in smaller tumors. However, a mixed phenotype showing the presence of both, glycolytic and gluconeogenic cancer cells was frequent. In lung adenocarcinoma, PCK2 expression was associated with significantly improved overall survival, while the opposite was found for GLUT1. The metabolic tumor microenvironment and the 3-dimensional context play an important role in modulating both pathways, since PCK2 expression preferentially occurred at the tumor margin and hypoxia regulated both, glycolysis and gluconeogenesis, in NSCLC cells in vitro, albeit in opposite directions. PCK1/2 expression was enhanced in metastases compared to primary tumors, possibly related to the different environment. The results of this study show that glycolysis and gluconeogenesis are activated in NSCLC in a tumor size and oxygenation modulated manner and differentially correlate with outcome. The frequent co-activation of gluconeogenesis and glycolysis in NSCLC should be considered in potential future therapeutic strategies targeting cancer cell metabolism.
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Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Gluconeogénesis , Glucólisis , Neoplasias Pulmonares/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Línea Celular Tumoral , Femenino , Transportador de Glucosa de Tipo 1/análisis , Transportador de Glucosa de Tipo 1/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Neoplasias Pulmonares/diagnóstico , Masculino , Fosfoenolpiruvato Carboxiquinasa (ATP)/análisis , Fosfoenolpiruvato Carboxiquinasa (ATP)/metabolismo , Fosfoenolpiruvato Carboxiquinasa (GTP)/análisis , Fosfoenolpiruvato Carboxiquinasa (GTP)/metabolismo , PronósticoRESUMEN
Importance: Identifying genes and proteins for cognitive resilience (ie, targets that may be associated with slowing or preventing cognitive decline regardless of the presence, number, or combination of common neuropathologic conditions) provides a complementary approach to developing novel therapeutics for the treatment and prevention of Alzheimer disease and related dementias. Objective: To identify proteins associated with cognitive resilience via a proteome-wide association study of the human dorsolateral prefrontal cortex. Design, Setting, and Participants: This study used data from 391 community-dwelling older persons who participated in the Religious Orders Study and the Rush Memory and Aging Project. The Religious Orders Study began enrollment January 1, 1994, and the Rush Memory and Aging Project began enrollment September 1, 1997, and data were collected and analyzed through October 23, 2019. Exposures: Participants had undergone annual detailed clinical examinations, postmortem evaluations, and tandem mass tag proteomics analyses. Main Outcomes and Measures: The outcome of cognitive resilience was defined as a longitudinal change in cognition over time after controlling for common age-related neuropathologic indices, including Alzheimer disease, Lewy bodies, transactive response DNA-binding protein 43, hippocampal sclerosis, infarcts, and vessel diseases. More than 8000 high abundance proteins were quantified from frozen dorsolateral prefrontal cortex tissue using tandem mass tag and liquid chromatography-mass spectrometry. Results: There were 391 participants (273 women); their mean (SD) age was 79.7 (6.7) years at baseline and 89.2 (6.5) years at death. Eight cortical proteins were identified in association with cognitive resilience: a higher level of NRN1 (estimate, 0.140; SE, 0.024; P = 7.35 × 10-9), ACTN4 (estimate, 0.321; SE, 0.065; P = 9.94 × 10-7), EPHX4 (estimate, 0.198; SE, 0.042; P = 2.13 × 10-6), RPH3A (estimate, 0.148; SE, 0.031; P = 2.58 × 10-6), SGTB (estimate, 0.211; SE, 0.045; P = 3.28 × 10-6), CPLX1 (estimate, 0.136; SE, 0.029; P = 4.06 × 10-6), and SH3GL1 (estimate, 0.179; SE, 0.039; P = 4.21 × 10-6) and a lower level of UBA1 (estimate, -0.366; SE, 0.076; P = 1.43 × 10-6) were associated with greater resilience. Conclusions and Relevance: These protein signals may represent novel targets for the maintenance of cognition in old age.
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Adaptación Psicológica , Disfunción Cognitiva/sangre , Vida Independiente/estadística & datos numéricos , Proteínas/análisis , Actinina/análisis , Actinina/sangre , Proteínas Adaptadoras Transductoras de Señales/análisis , Proteínas Adaptadoras Transductoras de Señales/sangre , Proteínas Adaptadoras del Transporte Vesicular/análisis , Proteínas Adaptadoras del Transporte Vesicular/sangre , Anciano , Anciano de 80 o más Años , Disfunción Cognitiva/epidemiología , Epóxido Hidrolasas/análisis , Epóxido Hidrolasas/sangre , Femenino , Proteínas Ligadas a GPI/análisis , Proteínas Ligadas a GPI/sangre , Humanos , Vida Independiente/psicología , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/sangre , Masculino , Chaperonas Moleculares/análisis , Chaperonas Moleculares/sangre , Proteínas del Tejido Nervioso/análisis , Proteínas del Tejido Nervioso/sangre , Neuropéptidos/análisis , Neuropéptidos/sangre , Enzimas Activadoras de Ubiquitina/análisis , Enzimas Activadoras de Ubiquitina/sangre , Proteínas de Transporte Vesicular/análisis , Proteínas de Transporte Vesicular/sangre , Rabfilina-3ARESUMEN
Scutellarein (SCU), a flavone that belongs to the flavonoid family and abundantly present in Scutellaria baicalensis a flowering plant in the family Lamiaceae, has been reported to exhibit anticancer effects in several cancer cell lines including gastric cancer (GC). Although our previous study documented the mechanisms of Scutellareininduced cytotoxic effects, the literature shows that the proteomic changes that are associated with the cellular response to SCU have been poorly understood. To avoid adverse sideeffects and significant toxicity of chemotherapy in patients who react poorly, biomarkers anticipating therapeutic responses are imperative. In the present study, we utilized a comparative proteomic analysis to identify proteins associated with Scutellarein (SCU)induced cell death in GC cells (AGS and SNU484), by integrating twodimensional gel electrophoresis (2DE), mass spectrometry (MS), and bioinformatics to analyze the proteins. Proteomic analysis between SCUtreated and DMSO (control) samples successfully identified 41 (AGS) and 31 (SNU484) proteins by MALDITOF/MS analysis and protein database search. Comparative proteomics analysis between AGS and SNU484 cells treated with SCU revealed a total of 7 protein identities commonly expressed and western blot analysis validated a subset of identified critical proteins, which were consistent with those of the 2DE outcome. Molecular docking studies also confirmed the binding affinity of SCU towards these critical proteins. Phosphatidylinositol 4,5bisphosphate 3kinase catalytic subunit ß isoform (PIK3CB) protein expression was accompanied by a distinct group of cellular functions, including cell growth, and proliferation. Cancerous inhibitor of protein phosphatase 2A (CIP2A), is one of the oncogenic molecules that have been shown to promote tumor growth and resistance to apoptosis and senescenceinducing therapies. In the present study, both PIK3CB and CIP2A proteins were downregulated in SCUtreated cells, which boosts our previous results of SCU to induce apoptosis and inhibits GC cell growth by regulating these critical proteins. The comparative proteomic analysis has yielded candidate biomarkers of response to SCU treatment in GC cell models and further validation of these biomarkers will help the future clinical development of SCU as a novel therapeutic drug.
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Antineoplásicos Fitogénicos/farmacología , Apigenina/farmacología , Biomarcadores de Tumor/genética , Neoplasias Gástricas/tratamiento farmacológico , Antineoplásicos Fitogénicos/uso terapéutico , Apigenina/uso terapéutico , Apoptosis/efectos de los fármacos , Apoptosis/genética , Autoantígenos/análisis , Autoantígenos/genética , Autoantígenos/metabolismo , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/genética , Fosfatidilinositol 3-Quinasa Clase I/análisis , Fosfatidilinositol 3-Quinasa Clase I/genética , Fosfatidilinositol 3-Quinasa Clase I/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Péptidos y Proteínas de Señalización Intracelular/análisis , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/análisis , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Simulación del Acoplamiento Molecular , Mapas de Interacción de Proteínas/efectos de los fármacos , Proteómica , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologíaRESUMEN
Endometrial cancer (EC) is the most common gynecologic malignancy in world. It has been reported that the mutation rate of FBXW7 is frequent in EC, but the specific functions of FBXW7 remain unknown in EC. In the present study, we revealed the role and mechanism of FBXW7 in EC cells. Compared with adjacent nontumor tissues, the FBXW7 expression level was lower in EC tissues. However, the level of STYX was in contrast with the expression of FBXW7 in EC tissues. And STYX interacted with FBXW7 and then down-regulated its expression level in EC. Over-expression of FBXW7 inhibited cell proliferation and facilitated apoptosis in EC cells, whereas silencing FBXW7 acted an opposite effect on EC cells. And the process of FBXW7 participated the proliferation and apoptosis in EC was regulated by STYX. FBXW7 suppressed the expression of Notch pathway related protein, and further inhibited the phosphorylation of mTOR. In addition, we also found that mTOR activitor (MHY1485) and Notch activator (Jagged-1) reversed the effect of over-expressing FBXW7 on cell proliferation and cell apoptosis. And Notch inhibitor (DAPT) counteracted the impact of over-expressing STYX on cell proliferation and cell apoptosis. Collectively, the present study verified that STYX inhibited the expression level of FBXW7 in EC, and then promoted cell proliferation but suppressed apoptosis through Notch-mTOR signaling pathway, which promoted carcinogenesis and progression of EC.