ABSTRACT
The identification of tumour biomarkers that detect the presence of disease using noninvasive diagnostic procedures is a key part of cancer research. We determined in plasma the vesicle-related microRNA (miRNA) expression profile of nonsmall cell lung cancer (NSCLC) and evaluate whether plasma miRNAs can be both discriminating (between patients and healthy controls) and prognostic markers. 365 human miRNAs were analysed by Taqman® low-density arrays (Applied Biosystems, Foster City, CA, USA) in the plasma from 28 NSCLC patients and 20 controls. Five selected miRNAs (let-7f, miR-20b, miR-30e-3p, miR-223 and miR-301) were validated independently by real-time PCR in plasma from 78 NSCLC and 48 controls and correlated with pathologic parameters and survival. Levels of let-7f, miR-20b and miR-30e-3p were decreased in plasma vesicles of NSCLC patients. Moreover, levels of let-7f and miR-30e-3p distinguished between two groups of patients for stage of disease and therefore possibility of surgery. Plasma levels of miR-30e-3p and let-7f were associated with short disease-free survival and overall survival, respectively. NSCLC patients and healthy controls differ in vesicle-related miRNAs in plasma. Levels of let-7f and miR-30e-3p in NSCLC patients are associated with poor outcome. Thus, plasma vesicle-related miRNAs obtained by noninvasive methods could serve as circulating tumour biomarkers of discriminating and prognostic value.
Subject(s)
Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/metabolism , MicroRNAs/metabolism , Biomarkers, Tumor , Carcinoma, Non-Small-Cell Lung/mortality , Case-Control Studies , Disease-Free Survival , False Positive Reactions , Female , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Genetic Markers , Humans , Lung Neoplasms/mortality , Male , PrognosisABSTRACT
Connexin (Cx) channels are sites of cytoplasmic communication between contacting cells. Evidence indicates that the opening of hemichannels occurs under both physiological and pathological conditions. In this paper, the involvement of Cx-43 hemichannels is demonstrated in the pathogenesis of Yersinia. Parental HeLa cells and transfected HeLa cells stably expressing Cx-43 (HCx43) were infected with Yersiniaenterocolitica, and bacterial uptake was measured by the colony-forming unit method. Bacterial uptake was higher in HCx43 cells than in parental cells and was inhibited by the Cx channel blocker, 18-alpha-glycyrrhetinic acid (AGA). The inhibitory effect of AGA was more pronounced on the Y. enterocolitica uptake by HCx43 cells than by parental cells. The ability of HCx43 cells to incorporate the permeable fluorescent tracer Lucifer Yellow (LY) was assessed. Dye incorporation was inhibited by AGA, whereas Y. enterocolitica infection of HCx43 cells increased LY incorporation. Western blotting analysis demonstrated that Y. enterocolitica infection of HCx43 cells induced tyrosine phosphorylation of Cx-43, thus supporting a critical role for Cx-43 in the strategies exploited by bacterial pathogens to invade non-phagocytic cells.
Subject(s)
Connexin 43/antagonists & inhibitors , Connexin 43/physiology , Glycyrrhetinic Acid/analogs & derivatives , Yersinia enterocolitica/pathogenicity , Animals , Bacterial Adhesion , Colony Count, Microbial , Glycyrrhetinic Acid/pharmacology , HeLa Cells , Humans , Ion Channel Gating/physiology , Isoquinolines/metabolism , Phosphorylation , Tyrosine/metabolismABSTRACT
Ciliary neurotrophic factor (CNTF) may be implicated in the pathogenetic mechanisms of hepatic encephalopathy. We tested this hypothesis by treating confluent primary cultures of rat astroglial cells with ammonium chloride for various periods and analysing the effect of ammonia on the signalling pathway that regulates CNTF mRNA and protein expression. Ammonia treatment induced a dose- and time-dependent reduction in CNTF mRNA and protein expression. Surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry analysis of CNTF in the culture medium demonstrated that ammonia also induced a significant decrease in CNTF release. In addition, ammonia affected Sp1 and c-fos, transcription factors that regulate CNTF mRNA and protein expression, which showed partial dephosphorylation and significantly lower mRNA and protein levels. Total content of p38MAPK (for which Sp1 and c-fos are substrates) was unaffected by ammonia, although the diphosphorylated (active) form was significantly reduced after ammonia exposure.
Subject(s)
Ammonia/pharmacology , Astrocytes/metabolism , Ciliary Neurotrophic Factor/biosynthesis , Genes, fos/drug effects , RNA, Messenger/biosynthesis , Signal Transduction/drug effects , Sp1 Transcription Factor/biosynthesis , p38 Mitogen-Activated Protein Kinases/biosynthesis , Animals , Astrocytes/drug effects , Blotting, Western , Cells, Cultured , Electrophoresis, Gel, Two-Dimensional , Phosphorylation , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Sp1 Transcription Factor/drug effects , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , p38 Mitogen-Activated Protein Kinases/drug effectsABSTRACT
In addition to the known compounds (+/-)-threo-guaiacylglycerol and the phenethyl alcohols, 3-methoxy-4-dihydroxyphenethyl alcohol and 3,4-dimethoxyphenethyl alcohol, a new irregular phenylpropanoid 2-(3-methoxy-4-hydroxyphenyl)-1,3-propanediol was isolated from the ethanolic extract of the leaves of Apollonias barbujana (Lauraceae). Their structures were elucidated on the basis of spectroscopic methods and chemical transformations.
