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1.
Anim Biotechnol ; 35(1): 2418516, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39460459

RESUMO

This study investigates whether Bacillus pumilus TS1 improves growth performance and alleviates inflammatory damage in broilers and explored its feasibility as an antibiotic alternative. We divided 240 one-day-old AA308 white-finned broilers into five groups (con, LPS, TS1L + LPS, TS1M + LPS and TS1H + LPS). The TS1L + LPS, TS1M + LPS and TS1H + LPS groups were fed TS1 for 15 days by gavage. The LPS, TS1L + LPS, TS1M + LPS and TS1H + LPS groups were injected intraperitoneally with 1 mg/kg LPS for three days. We investigated the probiotic and anti-inflammatory activities by measuring body weight, sequencing the intestinal flora and examining the structure of tissues by using pathological stain, real-time PCR, Western blotting and immunohistochemical detection. TS1 could improve growth performance and intestinal flora composition, also reduced different organ damage and inflammatory cytokine expression in serum and organs. The mechanism may involve upregulating HSP60 and HSP70 expression, targeting and regulating Nrf2 and P38 MAPK and modulating NF-κB and HO-1 expression at the transcriptional level in different organs. B. pumilus TS1 alleviated Inflammatory injury caused by LPS and attenuated the inflammatory response in broilers, and these effects were achieved through MAPK and Nrf2 regulation of HSPs/HO-1 in different organs. The above results suggested broilers fed with TS1 could release the LPS caused organ damage, and the most suggested dosage was 1.4 × 108 CFU/mL.


Assuntos
Anti-Inflamatórios , Bacillus pumilus , Galinhas , Microbioma Gastrointestinal , Inflamação , Lipopolissacarídeos , Probióticos , Animais , Bacillus pumilus/efeitos dos fármacos , Probióticos/farmacologia , Anti-Inflamatórios/farmacologia , Inflamação/tratamento farmacológico , Inflamação/induzido quimicamente , Microbioma Gastrointestinal/efeitos dos fármacos , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/induzido quimicamente , Citocinas/genética , Citocinas/metabolismo , Ração Animal/análise
2.
Int J Mol Sci ; 25(15)2024 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-39125776

RESUMO

Heat shock proteins (HSPs) play an important role in all living organisms under stress conditions by acting as molecular chaperones. The expression of different HSPs during stress varies depending on their protective functions and anti-apoptotic activities. The application of HSPs improves the efficiency and decreases the economic cost of animal breeding. By upregulating the expression of HSPs, feed supplements can improve stress tolerance in farm animals. In addition, high expression of HSPs is often a feature of tumor cells, and inhibiting the expression of HSPs is a promising novel method for killing these cells and treating cancers. In the present review, the findings of previous research on the application of HSPs in animal breeding and veterinary medicine are summarized, and the knowledge of the actions of HSPs in animals is briefly discussed.


Assuntos
Proteínas de Choque Térmico , Animais , Proteínas de Choque Térmico/metabolismo , Proteínas de Choque Térmico/genética , Cruzamento/métodos , Estresse Fisiológico , Resposta ao Choque Térmico
3.
BMC Vet Res ; 19(1): 41, 2023 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-36759839

RESUMO

BACKGROUND: In the current context of reduced and limited antibiotic use, several pathogens and stressors cause intestinal oxidative stress in poultry, which leads to a reduced feed intake, slow or stagnant growth and development, and even death, resulting in huge economic losses to the poultry breeding industry. Oxidative stress in animals is a non-specific injury for which no targeted drug therapy is available; however, the health of poultry can be improved by adding appropriate feed additives. Bacillus pumilus, as a feed additive, promotes growth and development and reduces intestinal oxidative stress damage in poultry. Heat shock protein 70 (HSP70) senses oxidative damage and repairs unfolded and misfolded proteins; its protective effect has been widely investigated. Mitogen-activated protein kinase/protein kinase C (MAPK/PKC) and hypoxia inducible factor-1 alpha (HIF-1α) are also common proteins associated with inflammatory response induced by several stressors, but there is limited research on these proteins in the context of poultry intestinal Salmonella Enteritidis (SE) infections. In the present study, we isolated a novel strain of Bacillus pumilus with excellent performance from the feces of healthy yaks, named TS1. To investigate the effect of TS1 on SE-induced enteritis in broilers, 120 6-day-old white-feathered broilers were randomly divided into four groups (con, TS1, SE, TS1 + SE). TS1 and TS1 + SE group chickens were fed with 1.4 × 107 colony-forming units per mL of TS1 for 15 days and intraperitoneally injected with SE to establish the oxidative stress model. Then, we investigated whether TS1 protects the intestine of SE-treated broiler chickens using inflammatory cytokine gene expression analysis, stress protein quantification, antioxidant quantification, and histopathological analysis. RESULTS: The TS1 + SE group showed lower MDA and higher GSH-Px, SOD, and T-AOC than the SE group. TS1 alleviated the effects of SE on intestinal villus length and crypt depth. Our results suggest that SE exposure increased the expression of inflammatory factors (IL-1ß, IL-6, TNF-α, IL-4, and MCP-1), p38 MAPK, and PKCß and decreased the expression of HSP60, HSP70, and HIF-1α, whereas TS1 alleviated these effects. CONCLUSIONS: Bacillus pumilus TS1 alleviated oxidative stress damage caused by SE and attenuated the inflammatory response in broilers through MAPK/PKC regulation of HSPs/HIF-1α.


Assuntos
Bacillus pumilus , Galinhas , Animais , Salmonella enteritidis , Intestinos , Mucosa Intestinal/metabolismo , Ração Animal/análise , Dieta/veterinária , Suplementos Nutricionais
4.
Mol Cell Proteomics ; 18(11): 2262-2272, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31501225

RESUMO

N-glycosylation alteration has been reported in liver diseases. Characterizing N-glycopeptides that correspond to N-glycan structure with specific site information enables better understanding of the molecular pathogenesis of liver damage and cancer. Here, unbiased quantification of N-glycopeptides of a cluster of serum glycoproteins with 40-55 kDa molecular weight (40-kDa band) was investigated in hepatitis B virus (HBV)-related liver diseases. We used an N-glycopeptide method based on 18O/16O C-terminal labeling to obtain 82 comparisons of serum from patients with HBV-related hepatocellular carcinoma (HCC) and liver cirrhosis (LC). Then, multiple reaction monitoring (MRM) was performed to quantify N-glycopeptide relative to the protein content, especially in the healthy donor-HBV-LC-HCC cascade. TPLTAN205ITK (H5N5S1F1) and (H5N4S2F1) corresponding to the glycopeptides of IgA2 were significantly elevated in serum from patients with HBV infection and even higher in HBV-related LC patients, as compared with healthy donor. In contrast, the two glycopeptides of IgA2 fell back down in HBV-related HCC patients. In addition, the variation in the abundance of two glycopeptides was not caused by its protein concentration. The altered N-glycopeptides might be part of a unique glycan signature indicating an IgA-mediated mechanism and providing potential diagnostic clues in HBV-related liver diseases.


Assuntos
Carcinoma Hepatocelular/diagnóstico , Glicopeptídeos/sangue , Glicoproteínas/sangue , Hepatite B/complicações , Imunoglobulina A/sangue , Neoplasias Hepáticas/diagnóstico , Proteoma/análise , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/virologia , Estudos de Casos e Controles , Glicosilação , Hepatite B/virologia , Vírus da Hepatite B/isolamento & purificação , Humanos , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/virologia , Polissacarídeos/metabolismo
5.
Pharm Dev Technol ; 25(7): 815-822, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32178565

RESUMO

Purpose: Voriconazole nanoparticles (API-NPs) were prepared by nanospray drying to improve the solubility of voriconazole and reduce its interindividual variability.Methods: The preparation procedure was optimized by central composite design-response surface methodology. The properties of the nanoparticles were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR) analyses. The solubility, dissolution, and stability of the API-NPs were determined experimentally. The pharmacokinetics were assessed based on rat plasma levels of voriconazole. An acute oral toxicity test of the API-NPs was performed in mice.Results: The powers were formulated using cetyltrimethylammonium chloride (CTAC) as the carrier material. SEM and particle size results showed that the API-NPs had a narrow particle size distribution. The XRD, DSC, and FTIR analyses show a decrease in crystallinity and a polymorphic transformation of the nanoparticles after nanospray drying. The solubility in water was approximately 15 times higher than that of voriconazole. The API-NP tablets exhibited significantly higher plasma exposure, namely, longer acting times and lower variability. The acute administration of voriconazole showed no toxic histopathological effects on organ tissue.Conclusion: The solubility of voriconazole was greatly improved, it showed higher bioavailability and safety, and the interindividual variability in voriconazole pharmacokinetics was reduced by nanospray drying.


Assuntos
Composição de Medicamentos/métodos , Nanopartículas/química , Nanopartículas/metabolismo , Voriconazol/síntese química , Voriconazol/farmacocinética , Animais , Antifúngicos/síntese química , Antifúngicos/farmacologia , Antifúngicos/toxicidade , Camundongos , Nanopartículas/toxicidade , Pós , Distribuição Aleatória , Ratos , Ratos Wistar , Testes de Toxicidade Aguda/métodos , Voriconazol/toxicidade , Difração de Raios X/métodos
6.
Clin Lab ; 64(1): 153-161, 2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-29479898

RESUMO

BACKGROUND: Recently, studies have reported that protein glycosylation plays an important role in the occurrence and development of cancer. Gastric cancer is a common cancer with high morbidity and mortality owing to most gastric cancers are discovered only at an advanced stage. Here, we aim to discover novel specific serum glycanbased biomarkers for gastric cancer. METHODS: A lectin microarray with 50 kinds of tumor-associated lectin was used to detect the glycan profiles of serum samples between early gastric cancer and healthy controls. Then lectin blot was performed to validate the differences. RESULTS: The result of the lectin microarray showed that the signal intensities of 13 lectins showed significant differences between the healthy controls and early gastric cancer. Compared to the healthy, the normalized fluorescent intensities of the lectins PWA, LEL, and STL were significantly increased, and it implied that their specifically recognized GlcNAc showed an especially elevated expression in early gastric cancer. Moreover, the binding affinity of the lectins EEL, RCA-II, RCA-I, VAL, DSA, PHA-L, UEA, and CAL were higher in the early gastric cancer than in healthy controls. These glycan structures containing GalNAc, terminal Galß 1-4 GlcNAc, Tri/tetraantennary N-glycan, ß-1, 6GlcNAc branching structure, α-linked fucose residues, and Tn antigen were elevated in gastric cancer. While the two lectins CFL GNL reduced their binding ability. In addition, their specifically recognized N-acetyl-D-galactosamine structure and (α-1,3) mannose residues were decreased in early gastric cancer. Furthermore, lectin blot results of LEL, STL, PHA-L, RCA-I were consistent with the results of the lectin microarray. CONCLUSIONS: The findings of our study clarify the specific alterations for glycosylation during the pathogenesis of gastric cancer. The specific high expression of GlcNAc structure may act as a potential early diagnostic marker for gastric cancer.


Assuntos
Detecção Precoce de Câncer/métodos , Lectinas/metabolismo , Análise Serial de Proteínas/métodos , Neoplasias Gástricas/metabolismo , Glicoproteínas/sangue , Glicosilação , Humanos , Polissacarídeos/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Neoplasias Gástricas/sangue , Neoplasias Gástricas/diagnóstico
7.
Future Oncol ; 13(23): 2053-2063, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28984474

RESUMO

AIM: We explored the expression of S100A6 and its role in intrahepatic cholangiocarcinoma (ICC). METHODS: The expression of S100A6 in ICC samples was detected by immunohistochemistry. In vitro experiments, we silenced and overexpressed S100A6 to investigate its role in cell functions. RESULTS: The expression of S100A6 was markedly increased in ICC tissues and cell lines. S100A6 overexpression was an independent risk factor for patients' survival. Silencing S100A6 resulted in a suppression of proliferation and p38/MAPK activity, while overexpressing S100A6 caused a promotion of proliferation and p38/MAPK. DISCUSSION:  S100A6 participated in the proliferation of ICC cells and correlated with a more aggressive behavior of ICC. Conclusion: S100A6 may serve as a novel prognostic marker and a potential therapeutic target for ICC patients.


Assuntos
Neoplasias dos Ductos Biliares/metabolismo , Proteínas de Ciclo Celular/metabolismo , Colangiocarcinoma/metabolismo , Sistema de Sinalização das MAP Quinases , Proteína A6 Ligante de Cálcio S100/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Adulto , Idoso , Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/mortalidade , Neoplasias dos Ductos Biliares/patologia , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Colangiocarcinoma/genética , Colangiocarcinoma/mortalidade , Colangiocarcinoma/patologia , Feminino , Seguimentos , Expressão Gênica , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Proteína A6 Ligante de Cálcio S100/genética
8.
Glycobiology ; 26(7): 684-692, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26873173

RESUMO

Protein glycosylation is one of the most significant post-translation modifications and plays a critical role in various biological functions. Haptoglobin (Hp) is one of the acute-phase response proteins secreted by liver. Its glycosylation could be analyzed by many analytical techniques qualitatively and quantitatively. The glycosylation alterations of Hp are reported to be associated with different kinds of diseases. The main glycosylation alterations of Hp in cancer appear to be the presence of aberrantly fucosylated and sialylated structures as well as increased branching. In this mini review, we provided a brief overview of Hp structure and biological function, discussed its glycosylation alterations in different cancers, and described the existing technologies for analyzing glycosylation site and glycan of Hp. Given the importance of Hp glycosylation, its unknown and unclear biological complexity and significances, Hp glycosylation has become a major target in cancer research. Development of sensitive and specific detection of Hp glycosylation including large-scale validation may be significant steps forward to its clinical application.


Assuntos
Carcinoma Hepatocelular/metabolismo , Haptoglobinas/metabolismo , Neoplasias Hepáticas/metabolismo , Fígado/metabolismo , Carcinoma Hepatocelular/patologia , Glicosilação , Haptoglobinas/genética , Humanos , Fígado/patologia , Neoplasias Hepáticas/patologia , Polissacarídeos/metabolismo , Processamento de Proteína Pós-Traducional/genética , Proteômica
9.
Int J Cancer ; 138(8): 1824-34, 2016 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-26853533

RESUMO

Heat shock proteins (HSPs) are highly conserved proteins, which are expressed at low levels under normal conditions, but significantly induced in response to cellular stresses. As molecular chaperones, HSPs play crucial roles in protein homeostasis, apoptosis, invasion and cellular signaling transduction. The induction of HSPs is an important part of heat shock response, which could help cancer cells to adapt to stress conditions. Because of the constant stress condition in tumor microenvironment, HSPs overexpression is widely reported in many human cancers. In light of the significance of HSPs for cancer cells to survive and obtain invasive phenotype under stress condition, HSPs are often associated with poor prognosis and treatment resistance in many types of human cancers. It has been described that upregulation of HSPs may serve as diagnostic and prognostic markers in hepatocellular carcinoma (HCC). Targeting HSPs with specific inhibitor alone or in combination with chemotherapy regimens holds promise for the improvement of outcomes for HCC patients. In this review, we summarize the expression profiles, functions and molecular mechanisms of HSPs (HSP27, HSP70 and HSP90) as well as a HSP-like protein (clusterin) in HCC. In addition, we address progression and challenges in targeting these HSPs as novel therapeutic strategies in HCC.


Assuntos
Carcinoma Hepatocelular/patologia , Proteínas de Choque Térmico/metabolismo , Neoplasias Hepáticas/patologia , Animais , Humanos
10.
Expert Rev Proteomics ; 13(9): 833-43, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27448621

RESUMO

INTRODUCTION: The liver is an important organ in humans. Hepatocellular carcinoma (HCC) is one of the deadliest cancers in the world. Progress in the Human Liver Proteome Project (HLPP) has improved understanding of the liver and the liver cancer proteome. AREAS COVERED: Here, we summarize the recent progress in liver proteome modification profiles, proteomic studies in liver cancer, proteomic study in the search for novel liver cancer biomarkers and drug targets, and progress of the Chromosome Centric Human Proteome Project (CHPP) in the past five years in the Institutes of Biomedical Sciences (IBS) of Fudan University. Expert commentary: Recent advances and findings discussed here provide great promise of improving the outcome of patients with liver cancer.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Fígado/metabolismo , Proteoma/genética , Carcinoma Hepatocelular/patologia , Humanos , Fígado/patologia , Neoplasias Hepáticas/patologia , Proteômica
12.
Proteomics ; 15(11): 1793-800, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25652264

RESUMO

Previously isolated pathways screened from individual genes were investigated at either the transcriptional or translational level; however, the consistency between the pathways screened at the gene expression levels was obscure in metastatic human hepatocellular carcinoma (HCC). To elucidate this question, we performed a transcriptomic (16,353 genes) and proteomic (7861 proteins) analysis simultaneously on six metastatic HCC cell lines against two nonmetastatic HCC cell lines, with all HBV traceable and close genetic-backgrounds for a comparative study. The quantitative and integrated results showed that significant genes were screened differentially with 351 transcripts from the transcriptome and 304 proteins from the proteome, with limited overlapping genes (7%). However, we discovered that these discrete 351 transcripts and 304 proteins screened share extrusive significant-pathways/networks with a 77% overlap, including active TGF-ß, RAS, NFκB, and Wnt, and inactive HNF4A, which are responsible for HCC metastasis. We conclude that the discrete, but significant genes predicted by either ome play intrinsically important roles in the linkage of responsible pathways shared by both omes in HCC metastasis.


Assuntos
Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Proteoma/análise , Carcinoma Hepatocelular/patologia , Transição Epitelial-Mesenquimal/genética , Perfilação da Expressão Gênica/métodos , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Neoplasias Hepáticas/patologia , Proteoma/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Via de Sinalização Wnt/genética , Proteínas ras/genética , Proteínas ras/metabolismo
13.
Cell Physiol Biochem ; 36(3): 1223-36, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26138883

RESUMO

BACKGROUND/AIMS: Anoikis resistance is a prerequisite for hepatocellular carcinoma (HCC) metastasis. The role of Caveolin-1 (CAV1) in anoikis resistance of HCC remains unclear. METHODS: The oncogenic effect of CAV1 on anchor-independent growth and anoikis resistance was investigated by overexpression and knockdown of CAV1 in hepatoma cells. IGF-1 pathway and its downstream signals were detected by immunoblot analysis. Caveolae invagination and IGF-1R internalization was studied by electron microscopy and (125)I-IGF1 internalization assay, respectively. The role of IGF-1R and tyrosine-14 residue (Y-14) of CAV1 was explored by deletion experiment and mutation experiment, respectively. The correlation of CAV1 and IGF-1R was further examined by immunochemical analysis in 120 HCC specimens. RESULTS: CAV1 could promote anchor-independent growth and anoikis resistance in hepatoma cells. CAV1-overexpression increased the expression of IGF-1R and subsequently activated PI3K/Akt and RAF/MEK/ERK pathway, while CAV1 knockdown showed the opposite effect. The mechanism study revealed that CAV1 facilitated caveolae invagination and (125)I-IGF1 internalization. IGF-1R deletion or Y-14 mutation reversed CAV1 mediated anchor-independent growth and anoikis resistance. In addition, CAV1 expression was positively related to IGF-1R expression in human HCC tissues. CONCLUSION: CAV1 confers resistance of hepatoma cells to anoikis by activating IGF-1 pathway, providing a potential therapeutic target for HCC metastasis.


Assuntos
Anoikis/genética , Carcinoma Hepatocelular/genética , Caveolina 1/genética , Regulação Neoplásica da Expressão Gênica , Fator de Crescimento Insulin-Like I/genética , Neoplasias Hepáticas/genética , Receptor IGF Tipo 1/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Cavéolas/metabolismo , Cavéolas/patologia , Caveolina 1/agonistas , Caveolina 1/antagonistas & inibidores , Caveolina 1/metabolismo , Adesão Celular , Linhagem Celular Tumoral , MAP Quinases Reguladas por Sinal Extracelular/genética , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , MAP Quinase Quinase Quinases/genética , MAP Quinase Quinase Quinases/metabolismo , Mutação , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Receptor IGF Tipo 1/metabolismo , Transdução de Sinais , Análise de Sobrevida , Quinases raf/genética , Quinases raf/metabolismo
14.
Glycoconj J ; 32(3-4): 119-25, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25702281

RESUMO

Serum paraoxonase 1 (PON1) is highly fucosylated in hepatocellular carcinoma (HCC) compared with liver cirrhosis (LC). Herein, lectin ELISA using Aleuria aurantia lectin (AAL) was established, which specifically measured optical density (OD) value of serum fucosylated PON1. PON1 protein ELISA was applied simultaneously. ELISA Index (OD value of fucosylated PON1/OD value of protein PON1) was introduced to indicate PON1 fucosylation level on its protein level (Fuc-PON1). ELISA Index in training group (90 LC and 90 HCC) was measured and area under the ROC curve (AUROC) was 0.803 with 80 % of sensitivity and 64.4 % of specificity in distinguishing early HCC from LC. Within training group, AFP(-) HCC (20/90) exhibited better AUROC (0.850), higher sensitivity (90 %) and specificity (75 %) than AFP(+) HCC (70/90). An independent testing set (20 LC and 20 HCC) validated the model and 17 HCC patients were successfully predicted. Meanwhile, serum AFP of 43 LC and 43 HCC had an AUROC of 0.760 with sensitivity of 79.1 % and specificity of 53.5 %. Thus, Fuc-PON1 may serve as a glycan biomarker for distinguishing early HCC from LC patients even with low AFP levels.


Assuntos
Arildialquilfosfatase/sangue , Carcinoma Hepatocelular/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Cirrose Hepática/diagnóstico , Neoplasias Hepáticas/diagnóstico , Idoso , Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/sangue , Feminino , Glicosilação , Humanos , Lectinas , Cirrose Hepática/sangue , Neoplasias Hepáticas/sangue , Masculino , Pessoa de Meia-Idade , Curva ROC , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , alfa-Fetoproteínas/análise
15.
Glycoconj J ; 32(9): 657-64, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26342810

RESUMO

Serum GP73 is a functional resident Golgi type II membrane protein with three potential N-glycosylation sites. In this study, we used multiple lectin assays to analyze glycan patterns of serum GP73 and evaluated its diagnostic value for distinguishing hepatocellular carcinoma (HCC) from liver cirrhosis (LC). Firstly, Antibody overlay lectin microarray and lectin blot were performed to observe altered glycans of GP73. Fucosylated structures were found to increase significantly in LC compared with HCC patients. Then, AAL ELISA assay using ELISA Index was utilized to measure fucosylation level of GP73 on its protein level (Fuc-GP73). ELISA Indices of 54 LC and 54 HCC patients was obtained and the area under the ROC curve (AUC) was 0.807 with a sensitivity of 85.2% and a specificity of 63.0% (cutoff of 3.182). In addition, combining Fuc-GP73 and AFP-L3 greatly improved the diagnostic accuracy (AUC = 0.953) and the diagnostic values were 94.4% sensitivity at 88.9% specificity. These data indicated that multiple lectin assays could contribute to pre-clinical evaluation of focused glycoprotein and Fuc-GP73 could act as a potential glycobiomarker complementary to AFP-L3 for discrimination of HCC from LC patients.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/sangue , Cirrose Hepática/sangue , Neoplasias Hepáticas/sangue , Proteínas de Membrana/sangue , Lectinas de Plantas/metabolismo , Processamento de Proteína Pós-Traducional , Idoso , Biomarcadores Tumorais/metabolismo , Feminino , Fucose/metabolismo , Glicosilação , Humanos , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Análise Serial de Proteínas , Ligação Proteica
16.
J Proteome Res ; 13(1): 126-36, 2014 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-24328083

RESUMO

We upgraded the preliminary CCPD 1.0 to CCPD 2.0 using the latest deep-profiling proteome (CCPD 2013) of three hepatocellular carcinoma (HCC) cell lines, namely, Hep3B, MHCC97H, and HCCLM3 (ProteomeXchange identifiers: PXD000529, PXD000533, and PXD000535). CCPD 2.0 totally covered 63.6% (438/689) of Chr. 8-coded proteins and 62.6% (439/701) of Chr. 8-coded protein-coding genes. Interestingly, we found that the missing proteins exhibited a tendency to form a cluster region in chromosomes, such as two ß-defensins clusters in Chr. 8, caused perhaps by their inflammation-related features. For the 41 Chr. 8-coded proteins being weakly or barely identified previously, we have performed an immunohistochemical (IHC) verification in 30 pairs of carcinoma/para-carcinoma HCC and 20 noncancerous liver tissues and confirmed their expressional evidence and occurrence proportions in tissue samples. We also verified 13 Chr. 8-coded HCC tumorigenesis-associated depleting or deficient proteins reported in CCPD 1.0 using IHC and screened 16 positive and 24 negative HCC metastatic potential-correlated proteins from large-scale label-free proteome quantitation data of CCPD 2013. Our results suggest that the selection of proper samples and the methodology to look for targeted missing proteins should be carefully considered in further verifications for the remaining Chr. 8-coded proteins.


Assuntos
Cromossomos Humanos Par 8 , Proteoma , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , China , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Biossíntese de Proteínas , Transcriptoma
17.
J Proteome Res ; 13(1): 38-49, 2014 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-24256510

RESUMO

To estimate the potential of the state-of-the-art proteomics technologies on full coverage of the encoding gene products, the Chinese Human Chromosome Proteome Consortium (CCPC) applied a multiomics strategy to systematically analyze the transciptome, translatome, and proteome of the same cultured hepatoma cells with varied metastatic potential qualitatively and quantitatively. The results provide a global view of gene expression profiles. The 9064 identified high confident proteins covered 50.2% of all gene products in the translatome. Those proteins with function of adhesion, development, reproduction, and so on are low abundant in transcriptome and translatome but absent in proteome. Taking the translatome as the background of protein expression, we found that the protein abundance plays a decisive role and hydrophobicity has a greater influence than molecular weight and isoelectric point on protein detectability. Thus, the enrichment strategy used for low-abundant transcription factors helped to identify missing proteins. In addition, those peptides with single amino acid polymorphisms played a significant role for the disease research, although they might negligibly contribute to new protein identification. The proteome raw and metadata of proteome were collected using the iProX submission system and submitted to ProteomeXchange (PXD000529, PXD000533, and PXD000535). All detailed information in this study can be accessed from the Chinese Chromosome-Centric Human Proteome Database.


Assuntos
Biossíntese de Proteínas , Proteoma , Transcriptoma , Linhagem Celular Tumoral , Perfilação da Expressão Gênica , Humanos , Espectrometria de Massas
18.
J Proteome Res ; 13(1): 114-25, 2014 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-24256544

RESUMO

Our first proteomic exploration of human chromosome 1 began in 2012 (CCPD 1.0), and the genome-wide characterization of the human proteome through public resources revealed that 32-39% of proteins on chromosome 1 remain unidentified. To characterize all of the missing proteins, we applied an OMICS-integrated analysis of three human liver cell lines (Hep3B, MHCC97H, and HCCLM3) using mRNA and ribosome nascent-chain complex-bound mRNA deep sequencing and proteome profiling, contributing mass spectrometric evidence of 60 additional chromosome 1 gene products. Integration of the annotation information from public databases revealed that 84.6% of genes on chromosome 1 had high-confidence protein evidence. Hierarchical analysis demonstrated that the remaining 320 missing genes were either experimentally or biologically explainable; 128 genes were found to be tissue-specific or rarely expressed in some tissues, whereas 91 proteins were uncharacterized mainly due to database annotation diversity, 89 were genes with low mRNA abundance or unsuitable protein properties, and 12 genes were identifiable theoretically because of a high abundance of mRNAs/RNC-mRNAs and the existence of proteotypic peptides. The relatively large contribution made by the identification of enriched transcription factors suggested specific enrichment of low-abundance protein classes, and SRM/MRM could capture high-priority missing proteins. Detailed analyses of the differentially expressed genes indicated that several gene families located on chromosome 1 may play critical roles in mediating hepatocellular carcinoma invasion and metastasis. All mass spectrometry proteomics data corresponding to our study were deposited in the ProteomeXchange under the identifiers PXD000529, PXD000533, and PXD000535.


Assuntos
Cromossomos Humanos Par 1 , Proteínas/genética , Linhagem Celular Tumoral , Humanos , Proteômica
19.
Int J Mol Sci ; 16(1): 178-92, 2014 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-25547487

RESUMO

The cAMP-regulated phosphoprotein 19 (ARPP-19) plays a key role in cell mitotic G2/M transition. Expression of ARPP-19 was increased in human hepatocellular carcinoma (HCC) compared to adjacent non-tumorous liver tissues in 36 paired liver samples, and the level of ARPP-19 in HCC tissues was positively correlated with the tumor size. To determine the interrelationship between ARPP-19 expression and HCC, we silenced ARPP-19 expression in the human hepatocarcinoma HepG2 and SMMC-7721 cells using lentivirus encoding ARPP-19 siRNA. HepG2 and SMMC-7721 cells with ARPP-19 knockdown displayed lowered cell growth rate, retarded colony formation and increased arrest at the G2/M phase transition. Silencing ARPP-19 in HCC cells resulted in decreased protein levels of phospho-(Ser) CDKs substrates and increased levels of inactivated cyclin division cycle 2 (Cdc2). Therefore, ARPP-19 may play a role in HCC pathogenesis through regulating cell proliferation.


Assuntos
Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Fosfoproteínas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteína Quinase CDC2 , Estudos de Casos e Controles , Proliferação de Células , Quinases Ciclina-Dependentes/genética , Quinases Ciclina-Dependentes/metabolismo , Feminino , Células Hep G2 , Humanos , Masculino , Pessoa de Meia-Idade , Fosfoproteínas/genética , Regulação para Cima
20.
Zhonghua Gan Zang Bing Za Zhi ; 22(9): 660-6, 2014 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-25369249

RESUMO

OBJECTIVE: To use a lectin microarray to study the alteration of glycan affinity profiles of serum glycoproteins during the hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) seroconversion in patients with chronic hepatitis B (CHB) following treatment with antiviral therapy,and to explore its biological significance. METHODS: CHB patients were divided into the following four groups:untreated HBeAg-positive,HBeAg seroconversion after anti-HBV therapy,HBsAg loss after anti-HBV therapy,and healthy individuals (controls).Serum samples were collected from each participant,depleted of high abundance proteins and analyzed by a lectin microarray containing 50 lectins.The lectin-affinity glycan profiles of serum proteins were partially verified by lectin blotting.Between-group differences were analyzed by one-way analysis of variance,and pairwise comparisons were carried out with the Student-Newman-Keuls (SNK) method. RESULTS: The results from the lectin microarray and lectin blotting assay showed significantly reduced affinity for 16 lectins in the untreated HBeAg-positive group compared to the control group (P less than 0.05);in addition,the specific glycan profiles of the untreated HBeAg-positive group included decreased terminal and core fructose,GalNAc alpha-Thr/Ser (T,Tn-antigen),GalNac alpha,terminal beta1-4,and beta-D galactose,bisecting and/or GlcNAc,mannose and Sia.However,the HBeAg seroconversion after anti-HBV therapy group showed enhanced binding of PSA,MPL and the above-mentioned 16 lectins (P less than 0.05),suggesting that the reduced serum glycoprotein glycan structures returned to normal or slightly higher than healthy levels after the therapy-induced seroconversion.Comparison of the group with HBsAg loss after anti-HBV therapy to the group with HBeAg seroconversion after anti-HBV therapy showed the binding ability of ten lectins (AAL,ACL,HAL,HPL,RCA-I,LEL,STL,PHA-E,NML and PCL) were weakened to near control levels and six lectins (VAL,LCA,GNL,PSA,MPL and JAC) were significantly strengthened (all P less than 0.05). These findings implied that the glycan containing terminal fructose, GalNacalpha, terminal beta1-4 galactose,and bisecting GlcNAc glycan structures dropped to near control levels, while the terminal beta-D-galactose residues and core fructose structure increased significantly. CONCLUSION: The glycan structures of serum glycoproteins are closely related to HBeAg and HBsAg seroconversion in CHB patients.It is possible that a special lectin binding glycan involving the terminal beta-D-galactose residues and core fructose may act as sugar markers associated with the disappearance of serum HBsAg during anti-HBV therapy for CHB.


Assuntos
Antivirais/uso terapêutico , Glicoproteínas/sangue , Hepatite B Crônica/tratamento farmacológico , Polissacarídeos/análise , Galactose , Antígenos E da Hepatite B , Humanos , Fito-Hemaglutininas
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