Project description:HCC cell lines with different metastatic potential (HepG2, Huh7, MHCC97 and PVTT cells) were treated with TGFbeta1 for 8 h We used microarrays to discover the long non-coding RNAs (lncRNAs) expression underlying Hepatocellular carcinoma (HCC) TGFbeta1 treated and non-treated control cells to identify distinct lncRNAs during this process.
Project description:4 replicates were prepared from A2058 melanoma cells [transfected with 10ng of empty vector (pcDNA3.1+)] and treated with 5ng/ml TGFβ1 or vehicle control for 24hrs This is the control arm of a larger experiment where cells transfected with a particular expression plasmid were treated with TGFβ1 or control vehicle. The transfection with the expresion plasmid was unsucessful so this empty vector data has been used alone to simply examine the effect of TGFβ1 treatment on A2058 cells.
Project description:To determine whether RSPO1 and TGFβ1 activate similar transcription, RNA-seq analysis was performed to compare gene profiles after treatment by RSPO1 or TGFβ1. FET cells were treated with RSPO1 or TGFβ1 in triplicates for 4 hours and RNAs extracted from these cells were used for RNA-seq analysis. When gene expression profiles were compared, it was found that 153 genes were commonly regulated by either RSPO1 or TGFβ1. Interestingly, the majority of genes regulated by TGFβ1 were also regulated by RSPO1.
Project description:To investigate TGFβ1 (a key member of the TGFβ superfamily) signaling in non-cancerous and cancerous cells derived from breast epithelium and what further response it might generate, we treated the cells with TGFβ1 or with conditioned media. We selected MCF10A non-tumorigenic breast epithelial cells (isolated from the mammary gland with fibrocystic disease) and MCF7 breast adenocarcinoma cells (estrogen receptor-positive, ER+). We followed a regimen of treating the cells for six consecutive days, with daily administration of TGFβ1 for two hours. The TGFβ1-containing medium was changed to a fresh one, which was collected after another 22 hours (so-called conditioned medium, CM) and administered to other cells. This enabled us to assess the ability of extracellularly secreted molecules to further induce a cellular response (bystander effect). RNA-seq analyses were performed on cells either directly treated with TGFβ1 in the above scheme or treated with a conditioned media for 1, 2, 3, 4, 5, or 6 days. The experiment was performed in three biological replicates, and each sample was sequenced separately. RNA-seg analysis in both cell lines allowed us to find the differences and similarities in response to TGFβ1 and CM. All 84 libraries were sequenced on Illumina NovaSeq6000 platform, in 2x150 mode, with a coverage of 60M reads/sample.
Project description:Cancer stem cells (CSCs) or tumor-initiating cells (TICs) organize a cellular hierarchy in a similar fashion to normal stem cell systems and exhibit high tumorigenic activity in xenograft transplantation assay. Disulfiram (DSF) could preferentially eradicate TICs, but the molecular machinery of its effect against TICs still remains largely unknown. We found that flow cytometric analyses showed that DSF but not 5-FU drastically reduces the number of tumor-initiating HCC cells. We conducted microarray analyses to examine gene expression profiling in DSF-treated tumor-initiating HCC cells. Purified EpCAM-positive HCC cells treated with Disulfiram or 5-FU were subjected to RNA extraction and hybridization on Agilent microarrays. Data were obtained for tripricate samples from three independent experiments.
Project description:4 replicates were prepared from A2058 melanoma cells [transfected with 10ng of empty vector (pcDNA3.1+)] and treated with 5ng/ml TGFβ1 or vehicle control for 24hrs