A search for genes modulated by interleukin-6 alone or with interleukin-1β in HepG2 cells using differential display analysis

AbstractInterleukin-1 and interleukin-6 are principal cytokines involved in regulation of expression of acute-phase proteins. In the joint action of both cytokines IL-1 can suppress or enhance the IL-6-dependent induction of gene expression. Here, we report changes in the transcriptome profile of HepG2 cells exposed to IL-6 alone, or IL-1 and IL-6. Cytokine-responsive genes were identified by differential display analysis. Validation of observed changes in the transcript level was carried out using the slot blot method. Out of 88 cDNA species modulated by IL-6, only 38 represent different known genes whereas 18 clones match genomic clones in NCBI data with hypothetical cDNA sequences (the remaining 32 clones showed no homology with the database or represented several clones of the same gene). In the experiments with HepG2 cells prestimulated for 3 h with IL-1 and then stimulated with IL-6, 43 cDNA fragments were amplified. Twenty-three of them represent known genes while 10 clones have inserts matching hypothetical cDNA sequences in NCBI data. The identified transcripts modulated by IL-6 or both cytokines in HepG2 cells code for intracellular proteins of various function. The largest groups represent genes engaged in metabolism, protein synthesis and signaling pathways. Among all genes identified as differentially regulated under stimulation by IL-6, or IL-1/IL-6, six were detected in both types of stimulation. None of the typical genes coding for plasma acute phase proteins was identified in our experiments. This indicates that differential display cannot be used to characterize the profile of a given transcriptome. On the other hand, it is a useful technique for detection of new genes responding to IL-6 alone or IL-6 in combination with IL-1

Thermophysical properties of IoNanofluids composed of 1-ethyl-3- methylimidazolium thiocyanate and carboxyl-functionalized long multi-walled carbon nanotubes

The concept of IoNanofluids (INFs) as the stable dispersions of nanoparticles in ionic liquids was proposed in 2009 by Nieto de Castro’s group. INFs characterize exciting properties such as improved thermal conductivity, non-volatility, and non-flammability. This work is a continuation of our studies on the morphology and physicochemistry of carbon-based nanomaterials a ecting thermal conductivity, viscosity, and density of INFs. We focus on the characterization of dispersions composed of long carboxylic group-functionalized multi-walled carbon nanotubes and 1-ethyl-3-methylimidazolium thiocyanate. The thermal conductivity of INFs was measured using KD2 Pro Thermal Properties Analyzer (Decagon Devices Inc., Pullman, WA, USA). The viscosity was investigated using rotary viscometer LV DV-II+Pro (Brookfield Engineering, Middleboro, MA, USA). The density of INFs was measured using a vibrating tube densimeter Anton Paar DMA 5000 (Graz, Austria). The maximum thermal conductivity enhancement of 22% was observed for INF composed of 1 wt% long carboxylic group-functionalized multi-walled carbon nanotubes

Identification of changes in the transcriptome profile of human hepatoma HepG2 cells stimulated with interleukin-1 beta

AbstractInterleukin-1 (IL-1) is the principal pro-inflammatory cytokine participating in the initiation of acute phase response. Human hepatoma HepG2 cells were exposed to 15 ng/ml of IL-1beta for times ranging from 1 to 24 h and the total RNA was isolated. Then cDNA was obtained and used for differential display with 10 arbitrary primers and 9 oligo(dT) primers designed by Clontech. Validation of observed changes of differentially expressed known genes was carried out by RT-PCR or Northern blot analysis. Out of 90 cDNA strands modulated by IL-1, 46 have been successfully reamplified and their sequencing indicates that they represent 36 different cDNA templates. By GenBank search, 26 cDNA clones were identified as already known genes while 10 showed no homology to any known gene. The identified transcripts modulated by IL-1 in HepG2 cells code for intracellular proteins of various function: trafficking/motor proteins (3 genes), proteins participating in the translation machinery or posttranscriptional/posttranslational modifications (7 genes), proteases (1 gene), proteins involved in metabolism (6 genes), activity modulators (3 genes), proteins of the cell cycle machinery (2 genes) and those functionally unclassified (4 genes). Majority of genes responded to IL-1 within 1 to 6 h (early genes), while two were late response genes (12–24 h) and four showed prolonged response over the whole 24-h period. Most of the observed changes of expression were in the range of two- to threefold increase in comparison to control untreated cells. Among identified genes, no typical secretory acute phase protein was found. The obtained results suggest that IL-1 affects the expression of several genes in HepG2 cells, especially those engaged in the synthesis and modifications of proteins