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1.
Genes (Basel) ; 11(1)2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31877923

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal cancer types world-wide. Its high mortality is related to the difficulty in the diagnosis, which often occurs when the disease is already advanced. As of today, no early diagnostic tests are available, while only a limited number of prognostic tests have reached clinical practice. The main reason is the lack of reliable biomarkers that are able to capture the early development or the progression of the disease. Hence, the discovery of biomarkers for early diagnosis or prognosis of PDAC remains, de facto, an unmet need. An increasing number of studies has shown that cell-free DNA (cfDNA) methylation analysis represents a promising non-invasive approach for the discovery of biomarkers with diagnostic or prognostic potential. In particular, cfDNA methylation could be utilized for the identification of disease-specific signatures in pre-neoplastic lesions or chronic pancreatitis (CP), representing a sensitive and non-invasive method of early diagnosis of PDAC. In this review, we will discuss the advantages and pitfalls of cfDNA methylation studies. Further, we will present the current advances in the discovery of pancreatic cancer biomarkers with early diagnostic or prognostic potential, focusing on pancreas-specific (e.g., CUX2 or REG1A) or abnormal (e.g., ADAMTS1 or BNC1) cfDNA methylation signatures in high risk pre-neoplastic conditions and PDAC.


Assuntos
Carcinoma Ductal Pancreático/genética , Ácidos Nucleicos Livres/genética , Proteína ADAMTS1/genética , Biomarcadores Tumorais/genética , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/metabolismo , Ácidos Nucleicos Livres/metabolismo , Metilação de DNA/genética , Proteínas de Ligação a DNA/genética , Epigênese Genética/genética , Proteínas de Homeodomínio/genética , Humanos , Litostatina/genética , Estadiamento de Neoplasias , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Pancreatite/genética , Pancreatite/metabolismo , Prognóstico , Regiões Promotoras Genéticas/genética , Fatores de Transcrição/genética
2.
Am J Physiol Cell Physiol ; 317(2): C366-C374, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31166710

RESUMO

Regenerating islet-derived (Reg) proteins, which were first discovered in the pancreas, are associated with increased proliferation, prevention of apoptosis, and enhanced differentiation in normal and disease states, but very little is known about the regulation of their expression. We hypothesized that Reg expression is influenced by microRNAs. Bioinformatic analysis predicted Reg1 to be a target of microRNA-7 (miR-7), which influences pancreatic ß-cell function. To this end, we investigated the effects of miR-7 on Reg1 expression in pancreatic acinar and islet ß-cells. High levels of Reg1 were noted by immunostaining and Western blotting in acinar cells in contrast to islet cells. A reciprocal expression pattern was observed for miR-7. Overexpression of miR-7 resulted in Reg1 mRNA suppression and reduction of secreted Reg1 protein. Conversely, miR-7 knockdown led to increases in Reg1. Targeting of Reg1 by miR-7 was confirmed via luciferase activity assays. In contrast, miR-7 did not directly repress the human ortholog of Reg1 REG1A as well as REG1B indicating species differences in the regulation of Reg expression. This is the first account of microRNA modulation of any Reg member warranting studies to fill gaps in our knowledge of Reg protein biology, particularly in disease contexts.


Assuntos
Células Secretoras de Insulina/metabolismo , Litostatina/metabolismo , MicroRNAs/metabolismo , Pâncreas Exócrino/metabolismo , Regiões 3' não Traduzidas , Animais , Sítios de Ligação , Linhagem Celular Tumoral , Regulação da Expressão Gênica , Células HEK293 , Humanos , Litostatina/genética , Camundongos Endogâmicos C57BL , MicroRNAs/genética , Pâncreas Exócrino/citologia , Especificidade da Espécie
3.
Biomed Res Int ; 2019: 6970890, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31139647

RESUMO

Pancreatic stone protein/regenerating protein Iα (PSP/REG Iα) is a secretory protein produced in the pancreas, but its expression has also been observed in the kidney. It may be associated with kidney dysfunction. This study investigates the possible association between PSP/REG Iα and kidney function in pregnant women. Serum PSP/REG Iα levels were measured by a specific ELISA enzyme-linked immunosorbent assay. Maternal information and clinical and biochemical parameters were collected. Estimated glomerular filtration rate (eGFR) was calculated for all individuals to evaluate their renal function. Spearman's correlation and multiple linear regression analyses were performed to assess the associations between PSP/REG Iα and eGFR, serum creatinine (Cr), blood urea nitrogen (BUN), and uric acid (UA). A total of 595 pregnant women were enrolled in the study. Participants with mildly reduced eGFR had higher PSP/REG Iα levels [50.49 (35.02, 58.64)] than in the general population [26.84 (21.02, 33.07)] (p < 0.001). Included participants were stratified into PSP/REG Iα quartiles; significant differences were observed in the levels of eGFR, serum Cr, BUN, and UA. PSP/REG Iα was negatively correlated with eGFR (r = -0.402, p < 0.001) and positively associated with serum Cr (r = 0.468, p < 0.001), BUN (r = 0.166, p < 0.001), and UA (r = 0.207, p < 0.001). The linear regression analysis indicated that PSP/REG Iα was associated with UA, BUN, and eGFR. High PSP/REG Iα concentrations were closely associated with renal dysfunction in pregnant women. Our study provides clinical evidence that serum PSP/REG Iα levels could be a novel biomarker for assessment of renal function in pregnant women.


Assuntos
Testes de Função Renal , Litostatina/sangue , Adulto , Feminino , Taxa de Filtração Glomerular , Humanos , Modelos Lineares , Gravidez
4.
Mol Cancer Res ; 17(7): 1459-1467, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30988165

RESUMO

Tumor protein p53 (TP53) mutation is a well-known occurrence at the late phase of carcinogenesis during the adenoma-carcinoma sequence of a sporadic colon cancer. Although numerous reports about clinical information of the patients with colon cancer have suggested that TP53 mutation might be related to various types of malignant potential, the direct effects of this mutation on the malignant potential of colon cancer remain unknown. Notably, no previous report has described a relationship between TP53 mutation and cancer stemness. We therefore aimed to assess the function of a TP53 mutant induced by the CRISPR-Cas9 system in colon cancer cells. In this study, two TP53 mutations, corresponding to exon 3 (TP53E3) and 10 (TP53E10), were generated in LS174T cells derived from a wild-type TP53 human colon cancer via a lentiviral CRISPR-Cas9 system. The loss of function of TP53 resulting from both mutations manifested as resistance to Nutlin3a-induced apoptosis and the downregulation of target genes of TP53. TP53 mutants exhibited an enhanced malignant potential, characterized by accelerated cell growth, invasiveness, chemoresistance, and cancer stemness. Interestingly, TP53E10 but not TP53E3 cells exhibited aberrant transcriptional activity of regenerating family member 1-α (REG1A) and expression of REG1A, resulting in the acquisition of enhanced malignant potential. In conclusion, we demonstrated for the first time that TP53 genomic mutation into human colon cancer cells affects the malignant potential. IMPLICATIONS: These findings suggest that both a loss of function and an aberrant gain of function of TP53 might promote high malignant potentials at the late phase of carcinogenesis in colon cancer.


Assuntos
Carcinogênese/genética , Neoplasias do Colo/genética , Litostatina/genética , Proteína Supressora de Tumor p53/genética , Sistemas CRISPR-Cas/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Resistencia a Medicamentos Antineoplásicos/genética , Éxons/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Mutação/genética , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia
5.
Biochim Biophys Acta Mol Basis Dis ; 1865(9): 2138-2148, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31029827

RESUMO

Defects in the development, maintenance or expansion of ß-cell mass can result in impaired glucose metabolism and diabetes. N6-methyladenosine affects mRNA stability and translation efficiency, and impacts cell differentiation and stress response. To determine if there is a role for m6A in ß-cells, we investigated the effect of Mettl14, a key component of the m6A methyltransferase complex, on ß-cell survival and function using rat insulin-2 promoter-Cre-mediated deletion of Mettl14 mouse line (ßKO). We found that ßKO mice with normal chow exhibited glucose intolerance, lower levels of glucose-stimulated insulin secretion, increased ß-cell death and decreased ß-cell mass. In addition, HFD-fed ßKO mice developed glucose intolerance, decreased ß-cell mass and proliferation, exhibited lower body weight, increased adipose tissue mass, and enhanced insulin sensitivity due to enhanced AKT signaling and decreased gluconeogenesis in the liver. HFD-fed ßKO mice also showed a decrease in de novo lipogenesis, and an increase in lipolysis in the liver. RNA sequencing in islets revealed that Mettl14 deficiency in ß-cells altered mRNA expression levels of some genes related to cell death and inflammation. Together, we showed that Mettl14 in ß-cells plays a key role in ß-cell survival, insulin secretion and glucose homeostasis.


Assuntos
Secreção de Insulina , Metiltransferases/metabolismo , Animais , Sobrevivência Celular , Dieta Hiperlipídica , Regulação da Expressão Gênica , Gluconeogênese , Intolerância à Glucose , Insulina/metabolismo , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/metabolismo , Lipogênese , Litostatina/genética , Litostatina/metabolismo , Fígado/metabolismo , Fígado/patologia , Masculino , Metiltransferases/antagonistas & inibidores , Metiltransferases/genética , Camundongos , Camundongos Knockout , Proteínas Associadas a Pancreatite/genética , Proteínas Associadas a Pancreatite/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo
6.
Cancer Biother Radiopharm ; 34(6): 362-370, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30973271

RESUMO

Objective: The reverse of chemoresistance and the improvement of sensitivity to chemotherapeutic agents of colorectal cancer cells have great clinical significance and the mechanism underlying the drug resistance is still unclear. REG Iα was reported to be upregulated in colorectal cancer tissues, but the roles of chemoresistance are still unclear. Materials and Methods: The expression of REG Iα in colorectal cancer cell lines was assessed by quantitative real-time polymerase chain reaction (Q-PCR). The expression of REG Iα in HCT116 and LOVO cells was knockdown by siRNA. The cell viability and IC50 (half maximal inhibitory concentration) values were analyzed by the CCK8 assay. The proportion of apoptosis and cell cycles were analyzed by flow cytometry. The migration potency of HCT116 and LOVO cells was analyzed by cell migration assay. The protein level of Cyclin D1, CDK4 (cyclin-dependent kinase 4), Bax and Bcl-2 were analyzed by western blot. Results: Knockdown of REG Iα enhances the sensitivity to 5-Fu of colorectal cancer cells. REG Iα knockdown promoted the cell apoptosis of HCT116 and LOVO under the 5-Fu treatment. The cell migration and cycle of colorectal cancer cells was also inhibited by REG Iα knockdown. We also found that REG Iα knockdown induced cell cycle arrest and cell apoptosis by Cyclin D1/CDK4 pathway and BAX/BCL-2 pathways. Conclusions: Knockdown of REG Iα enhances the sensitivity to 5-Fu of colorectal cancer cells via cyclin D1/CDK4 pathway and BAX/BCL-2 pathways.


Assuntos
Neoplasias Colorretais/tratamento farmacológico , Ciclina D1/metabolismo , Quinase 4 Dependente de Ciclina/metabolismo , Fluoruracila/farmacologia , Litostatina/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/metabolismo , Antimetabólitos Antineoplásicos/farmacologia , Apoptose , Ciclo Celular , Movimento Celular , Proliferação de Células , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Ciclina D1/genética , Quinase 4 Dependente de Ciclina/genética , Resistencia a Medicamentos Antineoplásicos , Regulação Neoplásica da Expressão Gênica , Humanos , Litostatina/genética , Litostatina/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Células Tumorais Cultivadas , Proteína X Associada a bcl-2/genética
7.
Biomark Med ; 13(2): 135-145, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30672312

RESUMO

Sepsis is a life-threatening syndrome characterized by a dysregulated host response to an infection resulting in multiple organ dysfunctions. Early diagnosis and management of sepsis is key to improve patient outcome but remains challenging. Despite extensive research, only few biomarkers have so far proven to be helpful in the diagnosis of sepsis. A novel protein biomarker, the pancreatic stone protein (PSP), is showing great promises. Several lines of evidences suggest that PSP has a higher diagnostic performance for the identification of sepsis than procalcitonin and C-reactive protein, and a strong prognostic value to predict unfavorable outcome at admission to intensive care unit. This review summarizes the current knowledge on the molecular mechanisms of PSP function and the clinical evidences available to highlight the relevance of this protein in the diagnosis and prognosis of sepsis.


Assuntos
Biomarcadores/análise , Biomarcadores/metabolismo , Litostatina/análise , Litostatina/metabolismo , Sepse/diagnóstico , Humanos , Prognóstico , Curva ROC , Sepse/metabolismo
8.
Biochem Genet ; 57(3): 382-402, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30600408

RESUMO

Regenerating islet-derived protein (Reg) could participate in the occurrence of diabetes mellitus, inflammation, tumors, and other diseased or damaged tissues. However, the correlation of Reg with acute hepatic failure (AHF) and hepatocellular carcinoma (HCC) is poorly defined. To reveal the expression profiles of Reg family and their possible regulatory roles in AHF and HCC, rat models of HCC and AHF were separately established, and Rat Genome 230 2.0 was used to detect expression profiles of Reg-mediated signaling pathways-associated genes from liver tissues in AHF and HCC. The results showed that a total of 79 genes were significantly changed. Among these genes, 67 genes were the AHF-specific genes, 45 genes were the HCC-specific genes, and 33 genes were the common genes. Then, K-means clustering classified these genes into 4 clusters based on the gene expression similarity, and DAVID analysis showed that the above altered genes were mainly associated with stress response, inflammatory response, and cell cycle regulation. Thereafter, IPA software was used to analyze potential effects of these genes, and the predicted results suggested that the Reg-mediated JAK/STAT, NF-κB, MAPK (ERK1/2, P38 and JNK), PLC, and PI3K/AKT signaling pathways may account for the activated inflammation and cell proliferation, and the attenuated apoptosis and cell death during the occurrence of AHF and HCC.


Assuntos
Carcinoma Hepatocelular/genética , Perfilação da Expressão Gênica , Litostatina/metabolismo , Falência Hepática Aguda/genética , Neoplasias Hepáticas/genética , Transdução de Sinais/genética , Animais , Carcinoma Hepatocelular/patologia , Ciclo Celular/genética , Modelos Animais de Doenças , Humanos , Inflamação/genética , Fígado/patologia , Falência Hepática Aguda/patologia , Neoplasias Hepáticas/patologia , Masculino , Família Multigênica , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estresse Fisiológico/genética
9.
Clin Chem Lab Med ; 57(4): 540-548, 2019 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-30240355

RESUMO

Background Early diagnosis of infection is essential for the initial management of cancer patients with chemotherapy-associated febrile neutropenia (FN). In this study, we have evaluated two emerging infection biomarkers, pancreatic stone protein (PSP) and soluble receptor of interleukin 2, known as soluble cluster of differentiation 25 (sCD25), for the detection of an infectious cause in FN, in comparison with other commonly used infection biomarkers, such as procalcitonin (PCT). Methods A total of 105 cancer patients presenting to the emergency department were prospectively enrolled. We observed 114 episodes of chemotherapy-associated FN. At presentation, a blood sample was collected for the measurement of PCT, PSP and sCD25. In order to evaluate the discriminatory ability of these markers for the diagnosis of infection, the area under the curve (AUC) of the receiver operating characteristic curves was calculated. Results Infection was documented in 59 FN episodes. PCT, PSP and sCD25 levels were significantly higher in infected patients. PCT was the biomarker with the highest diagnostic accuracy for infection (AUC: 0.901), whereas PSP and sCD25 showed a similar performance, with AUCs of 0.751 and 0.730, respectively. In a multivariable analysis, PCT and sCD25 were shown to be independently associated with infection. Conclusions Two novel biomarkers, PSP and sCD25, correlated with infection in cancer patients with chemotherapy-associated FN, but neither PSP nor sCD25 improved the performance of PCT. Based on the results obtained, the introduction of these novel biomarkers as a tool for the diagnosis of infection in this patient group is not recommended.


Assuntos
Neutropenia Febril/diagnóstico , Subunidade alfa de Receptor de Interleucina-2/sangue , Litostatina/sangue , Neoplasias/diagnóstico , Pró-Calcitonina/sangue , Idoso , Biomarcadores/sangue , Estudos de Coortes , Neutropenia Febril/sangue , Neutropenia Febril/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/sangue , Neoplasias/microbiologia , Estudos Prospectivos , Solubilidade
10.
Neurobiol Dis ; 119: 136-148, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30092268

RESUMO

REG-1α, a secreted protein containing a C-type lectin domain, is expressed in various organs and plays different roles in digestive system cells in physiological and pathological conditions. Like other members of the Reg family, REG-1α is expressed also in the brain where it has different functions. For instance, we previously reported that REG-1α regulates neurite outgrowth and is overexpressed during the very early stages of Alzheimer's disease (AD). However, REG-1α function in neural cells during neural degeneration remains unknown. First, REG-1α and phosphorylated tau expression were assessed in tissue sections from the hippocampus, representing neurofibrillary tangles (NFTs), from patients with AD, and from basal ganglia, representing subcortical NFTs, from patients with progressive supranuclear palsy (PSP). We found an association between REG-1α expression, tau hyperphosphorylation and NFTs in human brain samples from patients with these neurodegenerative diseases. Then, the effects of REG-1α overexpression on tau phosphorylation and axonal morphology were investigated i) in primary cultures of rat neurons that express human tau P301L and ii) in a transgenic zebrafish model of tauopathy that expresses human tau P301L. In the tau P301L cell model, REG-1α overexpression increased tau phosphorylation at the S202/T205 and S396 residues (early and late stages of abnormal phosphorylation, respectively) through the AKT/GSK3-ß pathway. This effect was associated with axonal defects both in tau P301L-expressing rat neurons and zebrafish embryos. Our findings suggest a functional role for REG-1α during tauopathy development and progression and, specifically, its involvement in the modification of tau phosphorylation temporal sequence.


Assuntos
Modelos Animais de Doenças , Litostatina/biossíntese , Tauopatias/metabolismo , Proteínas tau/biossíntese , Idoso , Idoso de 80 Anos ou mais , Animais , Animais Geneticamente Modificados , Encéfalo/metabolismo , Encéfalo/patologia , Células Cultivadas , Feminino , Humanos , Litostatina/genética , Masculino , Pessoa de Meia-Idade , Fosforilação/fisiologia , Ratos , Tauopatias/genética , Tauopatias/patologia , Peixe-Zebra , Proteínas tau/genética
11.
Biochem Biophys Res Commun ; 503(2): 963-969, 2018 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-29935186

RESUMO

Recently, we reported the presence of distinct cell clusters named acinar-like cell clusters touching Langerhans islets with thin interstitial surrounding (ATLANTIS) in human pancreas. A morphological study in humans demonstrated that ATLANTIS and islet cell clusters are found together in the microenvironment enclosed by a common basement membrane, and ATLANTIS releases vesicles containing Regenerating gene protein (REG Iα) to islet cell clusters. We examined 1) the presence or absence of ATLANTIS in homozygous Reg I (mouse homologue of human REG Iα) deficient (Reg I-/-) and wild-type mice, and 2) the possible role of ATLANTIS in the regeneration of beta cell clusters after encephalomyocarditis (EMC) virus (D-variant) infection in Reg I-/- and wild-type mice. ATLANTIS was found in both wild-type and Reg I-/- mice. In both groups, mean blood glucose increased transiently to greater than 14.0 mmol/L at 5 days after EMC virus infection and recovered to baseline at 12 days. At 12 days after EMC virus infection, lower BrdU labeling indices were observed in islet beta cells of Reg I-/- mice compared to wild-type mice. Beta cell volume 12 days after EMC virus infection in Reg I-/- mice did not differ from that of wild-type mice. These results suggest that Reg I, which is released from ATLANTIS to islet beta cell clusters, has a crucial role in beta cell regeneration in EMC virus-induced diabetes. The presence of mechanism(s) other than that mediated by Reg I in beta cell restoration after destruction by EMC virus was also suggested.


Assuntos
Infecções por Cardiovirus/complicações , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/virologia , Células Secretoras de Insulina/citologia , Litostatina/metabolismo , Pâncreas/citologia , Animais , Contagem de Células , Proliferação de Células , Células Cultivadas , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Vírus da Encefalomiocardite/isolamento & purificação , Deleção de Genes , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patologia , Células Secretoras de Insulina/virologia , Litostatina/genética , Masculino , Camundongos , Mitose , Pâncreas/metabolismo , Pâncreas/patologia , Pâncreas/virologia
12.
Sci Rep ; 8(1): 624, 2018 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-29330507

RESUMO

Compound Kushen Injection (CKI) is a Traditional Chinese Medicine (TCM) preparation that has been clinically used in China to treat various types of solid tumours. Although several studies have revealed that CKI can inhibit the proliferation of hepatocellular carcinoma (HCC) cell lines, the active compounds, potential targets and pathways involved in these effects have not been systematically investigated. Here, we proposed a novel idea of "main active compound-based network pharmacology" to explore the anti-cancer mechanism of CKI. Our results showed that CKI significantly suppressed the proliferation and migration of SMMC-7721 cells. Four main active compounds of CKI (matrine, oxymatrine, sophoridine and N-methylcytisine) were confirmed by the integration of ultra-performance liquid chromatography/mass spectrometry (UPLC-MS) with cell proliferation assays. The potential targets and pathways involved in the anti-HCC effects of CKI were predicted by a network pharmacology approach, and some of the crucial proteins and pathways were further validated by western blotting and metabolomics approaches. Our results indicated that CKI exerted anti-HCC effects via the key targets MMP2, MYC, CASP3, and REG1A and the key pathways of glycometabolism and amino acid metabolism. These results provide insights into the mechanism of CKI by combining quantitative analysis of components, network pharmacology and experimental validation.


Assuntos
Antineoplásicos Fitogênicos/análise , Carcinoma Hepatocelular/metabolismo , Medicamentos de Ervas Chinesas/análise , Neoplasias Hepáticas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Antineoplásicos Fitogênicos/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Caspase 3/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/farmacologia , Humanos , Litostatina/metabolismo , Neoplasias Hepáticas/tratamento farmacológico , Espectrometria de Massas , Metaloproteinase 2 da Matriz/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo
13.
Talanta ; 179: 472-477, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29310262

RESUMO

With the development of proteomics and the continuous discovery of biomarkers of trace proteins, it is important to accurately quantify low abundance protein, especially in urine for clinical diagnostics. In this paper, we reported a novel nano-biotinylated liposome-based immuno-loop-mediated isothermal amplification (LI-LAMP) for the ultrasensitive detection of REG1A (a biomarker for pancreatic ductal adenocarcinoma (PDAC) in urine) with high specificity. The detection range was 1µg/mL to 1fg/mL, with a detection limit of 1fg/mL, and no cross-reactivity was observed to occur in this assay. Compared with the amount of REG1A added, REG1A recovery using this method was 130% and 89%. Detection of REG1A concentrations using the LI-LAMP assay from real samples were in good agreement with those determined using ELISA, and relative deviations were not more than 10%. LI-LAMP shows good potential as a clinical diagnostic assay.


Assuntos
Biomarcadores Tumorais/urina , Carcinoma Ductal Pancreático/diagnóstico , Imunoensaio , Lipossomos/química , Litostatina/urina , Neoplasias Pancreáticas/diagnóstico , 1,2-Dipalmitoilfosfatidilcolina/análogos & derivados , 1,2-Dipalmitoilfosfatidilcolina/química , Biotinilação , Carcinoma Ductal Pancreático/patologia , Carcinoma Ductal Pancreático/urina , Colesterol/química , DNA/química , Humanos , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/urina , Reprodutibilidade dos Testes
14.
Med Sci Monit ; 23: 5834-5843, 2017 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-29222406

RESUMO

BACKGROUND Gastric cancer (GC) is one of the most common cause of cancer-related deaths. The clinical trials still lack the effective methods to treat or monitor the disease progression. In this research, the biological function and the underlying molecular mechanism of regenerating protein 1 alpha (REG1A) in GC were investigated. MATERIAL AND METHODS Gene expression omnibus (GEO), KMplot datasets and GC tissue microarray (n=164) were used to analyze the expression of REG1A and related patient prognoses in GC. Transwell matrigel assay, flow cytometry analysis and CCK8 cell viability assay were performed to detect the biological functions of REG1A. Western blotting and real-time PCR were used to detect the REG1A expression and PI3K/Akt related signaling. RESULTS It was found that the expression of REG1A was significantly downregulated in GC and closely related with clinicopathological findings or patient prognoses. REG1A overexpression could suppress the invasion, cell viability and promote the apoptosis of GC cells. Moreover, we found that the epigenetic methylation suppressed the expression level of REG1A in GC, and REG1A overexpression could suppress the phosphorylation of Akt or GSK3ß signaling. CONCLUSIONS Taken together, REG1A regulates cell invasion, apoptosis and viability in GC through activating PI3K/Akt-GSK3ß signaling. REG1A may serve as a promising therapeutic strategy for GC.


Assuntos
Litostatina/genética , Neoplasias Gástricas/genética , Apoptose/genética , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Sobrevivência Celular/genética , Metilação de DNA/genética , Regulação para Baixo , Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Litostatina/metabolismo , Prognóstico , Transdução de Sinais
15.
Med Sci Monit ; 23: 5211-5217, 2017 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-29091592

RESUMO

BACKGROUND High plasma levels of procalcitonin (PCT) are typically seen in children with severe bacterial infection, particularly in cases of septic shock or bacteremia. Similarly, pancreatic stone protein (PSP) is associated with inflammation, infection, and other disease-related stimuli. However, the prognostic value of PSP in critically ill pediatric patients is unknown. This study investigated the early diagnostic value of PCT and PSP in pediatric acute osteomyelitis. MATERIAL AND METHODS A total of 187 patients with suspected acute osteomyelitis and 80 healthy control children were enrolled. The serum expression of PTC and PSP was measured. Pearson correlation analysis was conducted to correlate PTC with PSP. ROC analysis was used to test the value of PTC and PSP in early diagnosis of pediatric acute osteomyelitis. RESULTS Acute osteomyelitis was diagnosed in 49.2% of the patients (n=92) based on the layered bone puncture. The serum levels of PTC and PSP in pediatric acute osteomyelitis were higher than in the non-acute osteomyelitis group (P<0.01). Serum PTC concentrations showed a significantly positive correlation with PSP levels (P<0.001). ROC analysis showed that the AUC values of PTC and PSP were 0.767 (95% CI, 0.700-0.826), and 0.796 (95% CI, 0.731-0.855), respectively. The AUC value of PTC & PSP was 0.903 (95% CI: 0.851-0.941), which was markedly increased compared with PTC or PSP (P<0.01). CONCLUSIONS Serum levels of PCT and PSP are promising biomarkers for early diagnosis of pediatric acute osteomyelitis.


Assuntos
Calcitonina/sangue , Litostatina/sangue , Osteomielite/sangue , Doença Aguda , Biomarcadores/sangue , Criança , Demografia , Feminino , Humanos , Modelos Logísticos , Masculino , Análise Multivariada , Osteomielite/diagnóstico , Curva ROC
16.
Biol Res ; 50(1): 37, 2017 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-29162157

RESUMO

BACKGROUND: Regenerating gene IA (REGIA) plays an important role in tissue regeneration and tumors prognosis of epithelium origin. However, the role of REGIA in nasopharyngeal carcinoma (NPC) is unclear. This study aims to investigate the expression and function of REG1A in NPC. RESULTS: We have found that there was 63 patients with REGIA positive expression of 155 patients in this study (40.65%). The positive expression rate of REGIA was 30.50, 44.44 and 47.83% in stage T2, T3 and T4 patients, respectively. The REGIA expression was significantly difference in T2 and T4 stage tumors or T2 and T3-T4 stage. The positive expression rate of REGIA was found to be higher in patients with cervical lymph node persistence than those with cervical lymph node complete regression. Patients with negative REGIA expression had a better overall survival and free survival than those with REGIA positive expression. In addition, according to the univariate and multivariate analysis, the REGIA expression was an independent adverse prognostic factor for NPC patients. CONCLUSION: REGIA expression was a useful biomarker in NPC patients for assessing T stage and survival.


Assuntos
Carcinoma/genética , Carcinoma/patologia , Litostatina/genética , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/análise , Biópsia , Carcinoma/mortalidade , Carcinoma/terapia , Progressão da Doença , Feminino , Humanos , Imuno-Histoquímica , Litostatina/fisiologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/mortalidade , Neoplasias Nasofaríngeas/terapia , Invasividade Neoplásica/patologia , Prognóstico , Estatísticas não Paramétricas
17.
Int J Mol Sci ; 18(11)2017 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-29077068

RESUMO

The periosteum is a thin membrane that surrounds the outer surface of bones and participates in fracture healing. However, the molecular signals that trigger/initiate the periosteal reaction are not well established. We fractured the rat femoral bone at the diaphysis and fixed it with an intramedullary inserted wire, and the expression of regenerating gene (Reg) I, which encodes a tissue regeneration/growth factor, was analyzed. Neither bone/marrow nor muscle showed RegI gene expression before or after the fracture. By contrast, the periosteum showed an elevated expression after the fracture, thereby confirming the localization of Reg I expression exclusively in the periosteum around the fractured areas. Expression of the Reg family increased after the fracture, followed by a decrease to basal levels by six weeks, when the fracture had almost healed. In vitro cultures of periosteal cells showed no Reg I expression, but the addition of IL-6 significantly induced Reg I gene expression. The addition of IL-6 also increased the cell number and reduced pro-apoptotic gene expression of Bim. The increased cell proliferation and reduction in Bim gene expression were abolished by transfection with Reg I siRNA, indicating that these IL-6-dependent effects require the Reg I gene expression. These results indicate the involvement of the IL-6/Reg pathway in the osteogenic response of the periosteum, which leads to fracture repair.


Assuntos
Consolidação da Fratura , Fraturas Ósseas/genética , Fraturas Ósseas/metabolismo , Regulação da Expressão Gênica , Interleucina-6/metabolismo , Litostatina/genética , Periósteo/metabolismo , Animais , Apoptose/genética , Proteína 11 Semelhante a Bcl-2/genética , Proteína 11 Semelhante a Bcl-2/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Fraturas Ósseas/patologia , Perfilação da Expressão Gênica , Imuno-Histoquímica , Litostatina/metabolismo , Masculino , Células-Tronco Mesenquimais/metabolismo , Periósteo/citologia , Periósteo/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos
18.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 48(3): 422-426, 2017 May.
Artigo em Chinês | MEDLINE | ID: mdl-28616918

RESUMO

OBJECTIVES: To determine the value of procalcitonin (PCT), high sensitivity C-reactive protein (hs-CRP) and pancreatic stone protein(PSP) in predicting the prognosis of children with sepsis. METHODS: A total of 106 hospitalized children [(4.4±1.6) year-old] with sepsis were enrolled in this study. The expressions of PTC, hs-CRP and PSP in the serum samples of the children were detected on the first day of admission to hospital. Pearson correlation analyses were performed to test the correlations between pediatric critical illness score (PCIS) and PTC, hs-CRP and PSP. Logistic regression models were established to determine factors predicting death of children. The value of PTC, hs-CRP and PSP in predicting the prognosis of children with sepsis was determined using ROC curves. RESULTS: About 32% children (34 cases) died. Higher expressions of PTC, hs-CRP and PSP were found in those who died (P<0.001). Serum PTC, hs-CRP and PSP were negatively correlated with PCIS (P<0.001). The multivariate logistic regression showed that PTC, hs-CRP and PSP were independent predictors of death in patients with sepsis (P<0.001). PTC, hs-CRP and PSP had an area under the curve (AUC) value of 0.86[ (95% confidence interval (CI), 0.78-0.92], 0.70 (95%CI, 0.61-0.79) and 0.69 (95%CI, 0.60-0.78) , respectively.The AUC value increased (P<0.001) to 0.92 (95%CI, 0.85-0.96) when the three indicators were combined (0.481×PCT+0.392×hs-CRP +0.314*PSP), with a value of less than 122.3 indicating good prognosis in 28 d. CONCLUSIONS: Serum PTC, hs-CRP and PSP can predict prognosis of children with sepsis.


Assuntos
Proteína C-Reativa/análise , Litostatina/sangue , Pró-Calcitonina/sangue , Sepse/diagnóstico , Biomarcadores/sangue , Criança , Pré-Escolar , Humanos , Prognóstico , Curva ROC
19.
Oncotarget ; 8(24): 38145-38151, 2017 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-28418911

RESUMO

Diabetic kidney disease (DKD) is a major complication of diabetes, and serves as an important cause of end-stage renal disease (ESRD). The role of chronic inflammation in DKD is becoming widely accepted. Pancreatic stone protein/regenerating protein (PSP/reg) is a secretory protein, which is elevated in blood during infected conditions and organ failure. The aim of this study was to investigate the relationship between serum PSP/reg and DKD in patients with type 2 diabetes (T2DM). A total of 120 subjects which includes newly diagnosed T2DM patients, diabetes patients without DKD, DKD patients, as well as healthy controls were enrolled in this study. Serum PSP/reg levels were significantly higher in DKD subjects compared with those of healthy controls (p < 0.001), newly diagnosed T2DM (p < 0.001) and diabetes patients without DKD (p < 0.001). PSP/reg levels correlated positively with glycated hemoglobin (HbA1c) (p < 0.001) and serum creatinine (p < 0.001). Meanwhile, serum PSP level was negatively correlated with estimated glomerular filtration rate (eGFR) (p < 0.001). The area under the curve (AUC) for presence of DKD was 0.854. IN CONCLUSION: PSP/reg levels are significantly up-regulated in DKD patients and might be related to renal injury. A follow-up study with a large cohort is needed.


Assuntos
Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/sangue , Litostatina/sangue , Adulto , Idoso , Área Sob a Curva , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Sensibilidade e Especificidade
20.
Oncotarget ; 8(18): 30162-30174, 2017 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-28415799

RESUMO

INTRODUCTION: In patients with infection and sepsis serum levels of Pancreatic Stone protein/regenerating protein I (PSP) are highly elevated. The origin of PSP during these conditions is presumably the pancreas, however, an intestinal origin cannot be excluded. Similarly, pancreatitis-associated protein (PAP) was identified in the pancreas. These proteins were also localized in intestinal organs. Here we aim to elucidate the bio-distribution of PSP and PAP in animal models of sepsis and in healthy humans. RESULTS: PSP and PAP responded to remote lesions in rats although the pancreatic response was much more pronounced than the intestinal. Tissue distribution of PSP demonstrated a 100-fold higher content in the pancreas compared to any other organ while PAP was most abundant in the small intestine. Both proteins responded to CLP or sham operation in the pancreas. PSP also increased in the intestine during CLP. The distribution of PSP and PAP in human tissue mirrored the distribution in the murine models. MATERIALS AND METHODS: Distribution of PSP and PAP was visualized by immunohistochemistry. Rats and mice underwent midline laparotomies followed by mobilization of tissue and incision of the pancreatic duct or duodenum. Standard cecum-ligation-puncture (CLP) procedures or sham laparotomies were performed. Human tissue extracts were analyzed for PSP and PAP. CONCLUSIONS: The pancreas reacts to remote lesions and septic insults in mice and rats with increased PSP synthesis, while PAP is selectively responsive to septic events. Furthermore, our results suggest that serum PSP in septic patients is predominantly derived through an acute phase response of the pancreas.


Assuntos
Litostatina/metabolismo , Pâncreas/metabolismo , Proteínas Associadas a Pancreatite/metabolismo , Estresse Fisiológico , Animais , Biomarcadores , Humanos , Masculino , Camundongos , Transporte Proteico , Ratos , Sepse/sangue , Sepse/etiologia , Sepse/metabolismo
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