ABSTRACT: Transcriptional profiling of human hepatocarcinoma comparing Huh-7 and SNU-739. Two-condition experiment, normalized ratio represented by Huh-7/SNU-739. Biological replicates: 2 Huh-7 replicates, 2 SNU-739 replicates.
Project description:MicroRNAs (miRNAs) are endogenously expressed single-stranded ~21–23 nucleotide RNAs that inhibit gene expression post-transcriptionally by binding imperfectly to elements usually within the 3′untranslated region (3′UTR) of mRNAs. Small interfering RNAs (siRNAs) mediate site-specific cleavage by binding with perfect complementarity to RNA. Here, a cell-based miRNA reporter system was developed to screen for compounds from marine and plant extracts that inhibit miRNA or siRNA activity. The daphnane diterpenoid genkwanine M (GENK) isolated from the plant Wikstroemia polyantha induces an early inflammatory response and can moderately inhibit miR-122 activity in the liver Huh-7 cell line. GENK does not alter miR-122 levels nor does it directly inhibit siRNA activity in an in vitro cleavage assay. Finally, we demonstrate that GENK can inhibit HCV infection in Huh-7 cells. In summary, the development of the cell-based miRNA sensor system should prove useful in identifying compounds that affect miRNA/siRNA activity. Control (no drug treatment): 1 hour and 4 hour samples, 3 replicates each; Drug-treated: 1 hour and 4 hours samples, 3 replicates each
Project description:mRNA expression profile modified by stable transfection of microRNA mir-517a (MIR517A) in a human hepatocellular carcinoma cell line Huh-7 Keywords: Hepatocellular carcinoma, Expression array, microRNA microRNA mir-517a (MIR517A) was transfected to Huh-7 cells using GFP-expressing lentiviral vector. Infected cells were selected using flow cytometry and subjected to mRNA expression microarray experiment. The profiles were compared to controls cells infected with only GFP protein.
Project description:A microarray time series was generated to identify which factor is expressed at different levels in two different gastric cancer cell lines, SNU 601 and SNU 668. These cell lines were treated for three different periods of time (24h, 48h, and 72h) with 1umol/L of olaparib or DMSO. Total RNA was extracted by using TRI reagent in accordance with the manufacturer`s instructions, and hybridized to an affymetrix GeneChip Human Gene 1.0 ST array. The results were normalized to the RObust Multi-Array Avarage (RMA) and analyzed.
Project description:Hepatitis C virus (HCV) infection is primarily treated with a pegylated interferon alpha based therapy, a regime that induces antiviral effects through the upregulation of many interferon-stimulated genes (ISGs). Whilst a number of anti-HCV ISGs have previously been identified, others may also be involved. Micorarrays were used to validate the presence of ISGs within subtracted libraries generated using the related techniques of suppression subtractive hybridisation and mirror orientation selection, which had initally been impllemented to isolate clones of ISGs following the interferon-alpha treatment of Huh-7 cells. Microarray data was generated for both untreated and interferon-alpha treated Huh-7 cells. No replicates were performed, however the microarray data was verified via the use of non-parametric (spearman) correlation analysis with RT-PCR data that had been generated using the same Huh-7 cell total RNA samples as the microarray experiments earlier.
Project description:TaqMan low density array (TLDA) was carried out to screen of the profiles of circulating miRNAs in pooled plasma samples from healthy controls and pre-operative osteosarcoma patients. The expression changes of circulating miRNAs in osteosarcoma patients were identified. To select candidate plasma miRNAs for osteosarcoma detection and monitoring, we employed TLDA technique to screen expression levels of 739 miRNAs in pooled plasma samples from healthy controls and pre-operative osteosarcoma patients (each pooled from 10 individuals).
Project description:To evaluate the effect on gene expression by shRNA-58335, we evaluated gene expression by microarray analysis. Gene expression was measured in Huh-7 cells stably expressing shRNA-58335 or control shRNA after infection with adenovirus expressing p53 (Ad-p53) or LacZ (Ad-LacZ).
Project description:To gain deeper insight into the mechanism of toxicity, it is important to identify and characterize mRNAs profiles involved in responses to specific classes of toxicants in conjunction with their impact on gene expression levels. However, few reports have described the effects of toxicants on mRNA expression profiles. Taking into account the prominent role of mRNAs in cancer development, progression, cell cycle control, and proliferation-related processes, it is likely that mRNAs are involved in the toxic response induced by carcinogens. Aldehydes are a well-characterized class of human carcinogens. In the present study, we documented the different expression profiles of mRNAs in environmental carcinogen-exposed A549 cells by mRNA microarray analysis. To evaluate the change in mRNA expression levels, human alveolar cells(A549) were exposed to seven lower-molecular-weight saturated aliphatic aldehyde (LSAAs) for 48h. mRNA expression analysis was conducted using a 4x44k human whole genome oligo microarray (Agilent Technologies, USA).
Project description:Analysis of Huh-7 hepatocarcinoma cell line depleted of NDRG3 or HIF-1α under hypoxic condition. HIF-1α and NDRG3 have distinct functions in hypoxia responses. Results provide insight into molecular basis of HIF-independent signaling in the development and progression of hypoxic tumors Gene expression profiles of Huh-7 cells stably expressing NDRG3-shRNA or HIF-1α-shRNA under normoxia were compared to gene expression profiles of Huh-7 stable cells under hypoxia for 3, 6, 12 and 24 hours.