SiRNA profiling of human colon cancer HT29 cells, siHX transfected vs. control
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ABSTRACT: HT29 cells were transsfected with siRNA (siHX), which is targeting the human endogeous retrovirus HERV-HX. Cells were harvested 72 h after transfection and knock-down of HERV-HX was evaluated by quantitative RT-PCR. Experiment Overall Design: Two-condition experiment, siHX vs. control cells. 1 transfected, 1 control.
Project description:HT29 cells were transsfected with siRNA (siHX), which is targeting the human endogeous retrovirus HERV-HX. Cells were harvested 72 h after transfection and knock-down of HERV-HX was evaluated by quantitative RT-PCR. Keywords: Genetic modification
Project description:HT29 cells was infected with EV71 at MOI 1 or nil respectively and harvested at 36hpi We use miRNA microarray to profile and identify miRNA which are up or down regulated due to EV71 infection 3 biological replicate (6 samples)
Project description:Our previous proteomics analysis suggested down-regulation of mitochondrial aconitase (ACO2) plays an important role in colorectal cancer. To evaluate the effect of mitochondrial aconitase overexpression on the metabolic and signaling pathway changes in colon cancer cell line HT29, we generated two ACO2 overexpressing cell lines #C and #UD and the mock-transfected vector control #VE. HT29 colon cancer cells overexpressing ACO2 exhibited lower rate of cell proliferation in vitro and reduced tumor growth potential in nude mice. In order to understand the signaling pathway changes, cDNA microarray analysis using GeneChips from Affymetrix were carried out.
Project description:The WWOX gene is a tumor suppressor probably involved in regulation of cell cycle and apoptosis and downregulated in variety of cancer types.However, its role in colon cancerogenesis is unknown. The aim of this study was to characterize how WWOX may be involved in colon cancerogenesis or cancer progression, how it influences the basic cancer cell features and modifies cell expression profile.Our observations suggest that in HT29 colon cancer cell line increased expression of WWOX may result in transition of cancer cells into more normal- like colon epithelium phenotype, on the other hand in SW480 WWOX revealed the well-known tumour suppressor properties. However, as the colon cancer is very heterogeneous disease, obtained discrepancies may reflect the known differences between cell lines and cancerogenesis pathway, which they undergone. HT29 colon cancer cells were stably transfected with WWOX cDNA. HT29 cells transfected with an empty vector served as a control. Total mRNA was isolated to look for gene-expression differences induced by the WWOX overexpression.
Project description:The experiment contains CUT&Tag profiles for transcription factor HNF4A in HT29 cell line treated with shRNA for PINK1. The experiment also include a control profile for HNF4A in HepG2 cell line.
Project description:TF antigen specific Sclerotium rolfsii lectin (SRL) induces apoptosis in human colon cancer HT29 cells and has tumour suppressing effect in vivo as reported earlier. In this study, we investigated the signaling pathways to identify the signature genes that lead to apoptosis of HT29 cells by mRNA and miRNA microarray analysis. HT29 cells were treated with SRL (20 ug/ml) for 2, 4, 8 12, 24 and 48h and gene expression microarray analysis was performed using SurePrint G3 Human Gene Expression Microarray kit. Several hundred entities were differentially regulated with a fold change value of greater than or equal to 2 and p value less than or equal to 0.05 following interaction of SRL at all-time points. Pathway analysis using GeneSpring 12.6.1 revealed that the MAPK signaling pathway is affected as early as 2h while cell cycle, DNA replication and apoptosis pathways are most significantly affected at 24 and 48h. SRL induced immediate over-expression of the transcription factor c-Jun as early as 2h. Very few miRNAs were found to be differentially regulated at 2 and 4h, however, a significant change in differentially expressed miRNAs were noticed at 12h with the miRNA gene target list significantly overlapping with the differential gene expression. The study suggests that the interaction of SRL with HT29 cells triggers apoptosis by affecting cell cycle, DNA replication and apoptosis signaling pathways. The observed effects may be initiated by affecting MAP kinase pathway and mediated by c-JUN. These findings were further validated by qRT-PCR and western blotting. The findings will enable to exploit and develop SRL as a possible targeted drug for cancer therapeutics.
Project description:Chromatin immunoprecipitation (ChIP-chip) study of histone 3 modifications (K4/K27) of 2 patients (colorectal cancer and normal tissue) and 1 cell line (HT29) on promoter arrays.
Project description:The representative cell line, HT29, culture in hypoxic and normoxic conditions were used to perform the miRNA microarray. The chip assay and data analysis were entrusted to KangChen Bio-tech, Inc. (Shanghai, China) and all the miRNAs with more than 2-fold variation in different samples were identified. Detailed methods were performed as previously described24 using the miRCURY Hy3 labeling kit and the microRNA array software (Exiqon, Denmark).