Project description:Oncogenic mutations in Kras initiate neoplastic transformation in the pancreas through a process that hijacks the activity of developmental regulators and induces an inflammatory microenvironment. We report that the homeodomain transcription factor Prox1 is a novel component of a progenitor signature that is activated in acinar cells undergoing dedifferentiation and ductal metaplasia conversion. Also, the conditional deletion of a single Prox1 allele markedly accelerates early transformation and significantly enhances features of inflammation in pancreatic tissues carrying a Kras oncogene. By using in vitro and in silico approaches, we demonstrate that Prox1 negatively regulates the expression of proinflammatory genes and genes implicated in malignant transformation in pancreatic cells. Microrrays were used to identify gene expression profiles and pathways that are differentially activated after enforced expression of PROX1 in the PROX1-negative human pancreatic ductal adenocarcinoma (PDAC) cell line Capan1. MSCV retroviral transduction was used to generate Capan1 cells that ectopically express PROX1 and selected for puromycin resistance. Three independent transductions with control virus (MSCV_puro) and 3 independent transductions with Prox1-cDNA virus (MSCV_PROX1_puro) were conducted for these experiments.
Project description:Glioblastoma represents the most common and aggressive primary brain tumor type in adults. Stem cell regulatory pathways have been shown to be activated in gliomas supporting self-renewal, tumor maintenance and survival under stress. Glioblastoma stem-like phenotype, cell motility and tumor cell heterogeneity are considered significant hurdles to overcome for developing new treatment against these tumors. Transcription factor PROX1 has been associated with stem-like-phenotypes. Here, we overexpressed and suppressed PROX1 in glioma cell lines in order understand the gene expression regulated by this transcription factor.
Project description:Analysis of CGTH-W-1 follicular thyroid carcinoma cells transcriptome following 48 hrs siRNA-mediated depletion of PROX1. PROX1 is a homeobox transcription factor. PROX1 depletion decreases migratory ability, motility and invasivness and induces profound cytoskeleton changes of CGTH-W-1 cells. Results provide insight into the role of PROX1 in the thyroid cancer. Three biological replicates for a given condition
Project description:Cell fate plasticity enables development, yet unlocked plasticity is a cancer hallmark. Regulating cell identity requires gene activation and repression. While master regulators induce lineage-specific genes to restrict plasticity, it remains unclear whether unwanted plasticity is actively suppressed by lineage-specific repressors. Here, we computationally predict so-called safeguard repressors for 18 cell types that block phenotypic plasticity lifelong. We validated hepatocyte-specific candidates using reprogramming, revealing that Prospero homeobox protein 1 (PROX1) enhanced hepatocyte identity by direct repression of alternate fate master regulators. In line with patient data, PROX1 overexpression in mice blocked initiation and progression of hepatocellular carcinoma and extended survival. Remarkably, Prox1 depletion caused hepatocyte fate loss in vitro, and promoted transdifferentiation of hepatocellular- to cholangio-carcinoma in vivo. Our findings provide mechanistic evidence for PROX1 as a hepatocyte-specific safeguard and support a model where individual cell type-specific repressors actively suppress plasticity throughout life to safeguard lineage choice and prevent disease.
Project description:To validate the suitability of two commonly used colorectal cancer cell lines, DLD1 and SW480, as model systems to study colorectal carcinogenesis, we treated these cell lines with beta-catenin siRNA and identified beta-catenin target genes using DNA microarrays. The list of identified target genes was compared to previously published beta-catenin target genes found in the PubMed and the GEO databases. Based on the large number of beta-catenin target genes found to be similarly regulated in DLD1, SW480 and LS174T as well as the large overlap with confirmed β-catenin target genes, we conclude that DLD1 and SW480 colon carcinoma cell lines are suitable model systems to study beta-catenin regulated genes and signaling pathways 12 arrays (2 cell lines, 2 treatments, 3 biological replicates)
Project description:Two commercial neuroblastoma cell lines (SIMA and SK-N-DZ) enriched their cancer stem cell (CSC) like cell population content by the use of conditioned cell culture media for neurospheres. Genome expression by microarray analysis was performed in CSC-like versus standard tumor cells culture.
Project description:Two isogenic human colorectal cancer cell lines (primary SW480 cell line and its lymph node metastatic variant SW620 cell line),as an in vitro metastatic model. We have demonstrated that SW620 cell line possesses high metastasis potential and SW480 cell linepossesses low metastatic potential. We want to compare the whole cell microRNAs profiles of two isogenic colorectal cancer cell lines (SW480 and SW620 cell line), to gain an insight into the molecular events of colon cancer metastasis.
Project description:RNA profiling of mouse haemopoetic lineages RNA profiling of mouse haemopoetic lineages. For the isolation and sequencing of ouse granulocyte nuclei, bone marrow from C57BL/6J was harvested from the femur, tibia and spine using a mortar and pestle in PBS supplemented with 2% (v/v) FCS and mature granulocytes purified by flow cytometry as described previously. Purification was validated by reanalysis by flow cytometry and May-GrM-CM-<nwald Giemsa staining. Nuclear purification was carried out using the PARIS kit (Ambion). RNA was extracted from the nuclear fraction using Trizol before deep sequencing. Deep sequencing was performed by GeneWorks (Adelaide, Australia) on the Illumina GAII according to the manufacturer's instructions. For more details, see Supplementary Methods at the NSMB website. Additional files and resources can be found at http://matticklab.com/index.php?title=NuclearTinyRNAs
Project description:Omega-3 polyunsaturated fatty acids are normal constituents of the diet and have an essential role in maintaining important cellular functions. Docosahexaenoic acid (DHA) have demonstrated anticancer activities in several in vitro and in vivo studies, and in some clinical studies. The mechanism by which n-3 PUFAs reduce tumor growth probably involves the inhibition of cell proliferation, induction of cell death, or a combination of both. There are differences in sensitivity towards DHA treatment among colorectal cell lines, although the reason why is unclear. 10 human colorectal cell lines, representing 5 different subtypes of colorectal cancer, were included in gene expression analysis. (Pleae refer to Sadanandam A, Lyssiotis CA, Homicsko K, Collisson EA, Gibb WJ, Wullschleger S, Ostos LC, Lannon WA, Grotzinger C, Del Rio M, et al. “A colorectal cancer classification system that associates cellular phenotype and responses to therapy”. Nat Med. 2013;19:619-625. doi:10.1038/nm.3175 for details on cancer subtype classifications.) Our aim is to identify different levels of genes and pathways correlating with differences in sensitivity towards DHA between the colorectal cell lines for future classification of patients that could benefit from omega-3 PUFA treatment in addition to conventional treatment.