Project description:L1 elements are repeat elements known to get activated during hepatocarcinogenesis. We have knocked-down expression of L1 elements using shRNA targeting L1-ORF1 in Huh7 cell line and a control non-targeting cell line was also generated. Influence of L1 knockdown on overall cellular transcritome was then analysed by RNAseq followed by pathway analysis. Overall, a crosstalk between L1 elements and TGFbeta signaling pathway is observed.
Project description:We use lentivirus carrying shRNA for THADA to evaluate the efect of THADA knockdown in Huh7 cells. The pGIPZ shRNA plasmid specific to human THADA (clone ID V3LHS_318037 Open Biosystems, Inc.) or the pGIPZ non-silencing shRNAmir lentiviral plasmid (as a control) was cotransfected with the pPACKH1 plasmid mix in HEK 293 cells. Stable clones of Huh7 cells with THADA knockdown were routinely maintained in a medium containing of puromycin (500ng/ml)
Project description:To identify CD24 signaling pathway Affymetrix Human Genome U133 Plus GeneChip 2.0 shRNA CD24 (knockdown) or non-target control (NTC) was stably transduced into Huh7 HCC cells by lentiviral approach
Project description:Analysis of Huh7 hepatocellular carcinoma (HCC) cells depleted for potassium channel KCa3.1, the intermediate conductance calcium-activated potassium channel. Result provide insight into the role of KCa3.1 in the carcinogenesis of HCC. We used microarrays to detail the gene expression differences between KCa3.1 knockdown and negative control in Huh7 HCC cells.
Project description:To identify candidate genes regulated by CD47 Affymetrix Human Genome U133 Plus GeneChip 2.0 HCC cell line Huh7 cells with or without CD47 repressed
Project description:The Wnt signaling pathway is involved in many differentiation events during embryonic development and can lead to tumor formation after aberrant activation of its components. Β-catenin, a cytoplasmic component, plays a major role in the transduction of the canonical wnt/ β-catenin signaling. The aim of this study was to identify novel genes that are regulated by active β-catenin/TCF signaling in hepatocellular carcinoma. We selected and expanded isogenic clones from hepatocellular carcinoma-derived Huh7 cells with high and low β-catenin/TCF activities. We showed that, high TCF activity Huh7 cells lead to bigger and more aggressive tumors when xenografted into nude mice. We used SAGE (Serial Analysis of Gene Expression), genome-wide microarray and in silico promoter analysis in parallel, to compare gene expression between low (basal) and high (transfected) β-catenin/TCF activity clones, those had been xenografted into nude mice. We compared and contrasted SAGE and genome-wide microarray data, in parallel. Finally; after combined analysis, we identified BRI3 and HSF2 as novel targets of Wnt/β-catenin signaling in hepatocellular carcinoma. Experiment Overall Design: High TCF activity Huh7 cell line (Huh7-S33Y) was compared to control Huh7 cell line (Huh7-Vec) by using 10 ug of total RNA isolated from each sample (15 ug of labeled cRNA was hybridized to the arrays). Triplicates are coming from same total RNA extraction.