Label-free quantitative proteomics of CD133-positive liver cancer stem cells

Abstract Background CD133-positive liver cancer stem cells, which are characterized by their resistance to conventional chemotherapy and their tumor initiation ability at limited dilutions, have been recognized as a critical target in liver cancer therapeutics. In the current work, we developed a label-free quantitative method to investigate the proteome of CD133-positive liver cancer stem cells for the purpose of identifying unique biomarkers that can be utilized for targeting liver cancer stem cells. Label-free quantitation was performed in combination with ID-based Elution time Alignment by Linear regression Quantitation (IDEAL-Q) and MaxQuant. Results Initially, IDEAL-Q analysis revealed that 151 proteins were differentially expressed in the CD133-positive hepatoma cells when compared with CD133-negative cells. We then analyzed these 151 differentially expressed proteins by MaxQuant software and identified 10 significantly up-regulated proteins. The results were further validated by RT-PCR, western blot, flow cytometry or immunofluorescent staining which revealed that prominin-1, annexin A1, annexin A3, transgelin, creatine kinase B, vimentin, and EpCAM were indeed highly expressed in the CD133-positive hepatoma cells. Conclusions These findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.http://deepblue.lib.umich.edu/bitstream/2027.42/113089/1/12953_2012_Article_407.pd

miRTarBase: a database curates experimentally validated microRNA–target interactions

MicroRNAs (miRNAs), i.e. small non-coding RNA molecules (∼22 nt), can bind to one or more target sites on a gene transcript to negatively regulate protein expression, subsequently controlling many cellular mechanisms. A current and curated collection of miRNA–target interactions (MTIs) with experimental support is essential to thoroughly elucidating miRNA functions under different conditions and in different species. As a database, miRTarBase has accumulated more than 3500 MTIs by manually surveying pertinent literature after data mining of the text systematically to filter research articles related to functional studies of miRNAs. Generally, the collected MTIs are validated experimentally by reporter assays, western blot, or microarray experiments with overexpression or knockdown of miRNAs. miRTarBase curates 3576 experimentally verified MTIs between 657 miRNAs and 2297 target genes among 17 species. miRTarBase contains the largest amount of validated MTIs by comparing with other similar, previously developed databases. The MTIs collected in the miRTarBase can also provide a large amount of positive samples to develop computational methods capable of identifying miRNA–target interactions. miRTarBase is now available on http://miRTarBase.mbc.nctu.edu.tw/, and is updated frequently by continuously surveying research articles

SJS/TEN 2019: From science to translation.

Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) are potentially life-threatening, immune-mediated adverse reactions characterized by widespread erythema, epidermal necrosis, and detachment of skin and mucosa. Efforts to grow and develop functional international collaborations and a multidisciplinary interactive network focusing on SJS/TEN as an uncommon but high burden disease will be necessary to improve efforts in prevention, early diagnosis and improved acute and long-term management. SJS/TEN 2019: From Science to Translation was a 1.5-day scientific program held April 26-27, 2019, in Vancouver, Canada. The meeting successfully engaged clinicians, researchers, and patients and conducted many productive discussions on research and patient care needs

Label-free quantitative proteomics of CD133-positive liver cancer stem cells

Abstract Background CD133-positive liver cancer stem cells, which are characterized by their resistance to conventional chemotherapy and their tumor initiation ability at limited dilutions, have been recognized as a critical target in liver cancer therapeutics. In the current work, we developed a label-free quantitative method to investigate the proteome of CD133-positive liver cancer stem cells for the purpose of identifying unique biomarkers that can be utilized for targeting liver cancer stem cells. Label-free quantitation was performed in combination with ID-based Elution time Alignment by Linear regression Quantitation (IDEAL-Q) and MaxQuant. Results Initially, IDEAL-Q analysis revealed that 151 proteins were differentially expressed in the CD133-positive hepatoma cells when compared with CD133-negative cells. We then analyzed these 151 differentially expressed proteins by MaxQuant software and identified 10 significantly up-regulated proteins. The results were further validated by RT-PCR, western blot, flow cytometry or immunofluorescent staining which revealed that prominin-1, annexin A1, annexin A3, transgelin, creatine kinase B, vimentin, and EpCAM were indeed highly expressed in the CD133-positive hepatoma cells. Conclusions These findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.</p

Electrochemical Polymerization of PEDOT–Graphene Oxide–Heparin Composite Coating for Anti-Fouling and Anti-Clotting of Cardiovascular Stents

In this study, a novel hemocompatible coating on stainless steel substrates was prepared by electrochemically copolymerizing 3,4-ethylenedioxythiophene (EDOT) with graphene oxide (GO), polystyrene sulfonate (PSS), or heparin (HEP) on SUS316L stainless steel, producing an anti-fouling (anti-protein adsorption and anti-platelet adhesion) surface to avoid the restenosis of blood vessels. The negative charges of GO, PSS, and HEP repel negatively charged proteins and platelets to achieve anti-fouling and anti-clotting. The results show that the anti-fouling capability of the poly(3,4-ethylenedioxythiophene) (PEDOT)/PSS coating is similar to that of the PEDOT/HEP coating. The anti-fouling capability of PEDOT/GO is higher than those of PEDOT/HEP and PEDOT/PSS. The reason for this is that GO exhibits negatively charged functional groups (COO&minus;). The highest anti-fouling capability was found with the PEDOT/GO/HEP coating, indicating that electrochemical copolymerization of PEDOT with GO and HEP enhances the anti-fouling capability. Furthermore, the biocompatibility of the PEDOT coatings was tested with 3T3 cells for 1&ndash;5 days. The results show that all PEDOT composite coatings exhibited biocompatibility. The blood clotting time (APTT) of PEDOT/GO/HEP was prolonged to 225 s, much longer than the 40 s of pristine SUS316L stainless steel (the control), thus greatly improving the anti-blood-clotting capability of cardiovascular stents

Flux balance analysis predicts Warburg-like effects of mouse hepatocyte deficient in miR-122a

<div><p>The liver is a vital organ involving in various major metabolic functions in human body. MicroRNA-122 (miR-122) plays an important role in the regulation of liver metabolism, but its intrinsic physiological functions require further clarification. This study integrated the genome-scale metabolic model of hepatocytes and mouse experimental data with germline deletion of <i>Mir122a</i> (<i>Mir122a</i>^{<i>–/–</i>}) to infer Warburg-like effects. Elevated expression of <i>MiR-122a</i> target genes in <i>Mir122a</i>^{<i>–/–</i>}mice, especially those encoding for metabolic enzymes, was applied to analyze the flux distributions of the genome-scale metabolic model in normal and deficient states. By definition of the similarity ratio, we compared the flux fold change of the genome-scale metabolic model computational results and metabolomic profiling data measured through a liquid-chromatography with mass spectrometer, respectively, for hepatocytes of 2-month-old mice in normal and deficient states. The <i>Ddc</i> gene demonstrated the highest similarity ratio of 95% to the biological hypothesis of the Warburg effect, and similarity of 75% to the experimental observation. We also used 2, 6, and 11 months of mir-122 knockout mice liver cell to examined the expression pattern of DDC in the knockout mice livers to show upregulated profiles of DDC from the data. Furthermore, through a bioinformatics (LINCS program) prediction, BTK inhibitors and withaferin A could downregulate DDC expression, suggesting that such drugs could potentially alter the early events of metabolomics of liver cancer cells.</p></div