Transcriptomic profiling of CDCP1 knock-down in human cardiac fibroblasts (HCFs)
Ontology highlight
ABSTRACT: To investigate the function of CDCP1 in regulation of HCF proliferation, we cultured primary HCFs and transfected with non-targeting siRNA and CDCP1 siRNAs. Total RNA samples were extracted for RNA-seq to identify transcriptome-wide gene expression changes in the HCFs after CDCP1 knock-down (KD).
Project description:Analysis of MDCK (type I) cysts overexpressing CDCP1-EGFP or its mutant. After cystogenesis in collagen matrix, overexpression of exogenous CDCP1 proteins was induced by the Tet-On system.
Project description:This experiment aims at comparing the transcriptomic profiles of two subpopulations of fetal human cardiac fibroblasts (HCF) defined by their relative expression of VCAM1. VCAM1-positive and VCAM1-negative populations were isolated from commercial fetal HCF by MACS against VCAM1. The RNA-seq procedure was outsourced to GENEWIZ (South Plainfield, NJ, USA), and subsequent analysis was performed in-house. Briefly, total mRNA was captured using the NEBNext Poly(A) mRNA Magnetic Isolation Module, and the library was constructed with the NEBNext Ultra RNA Library Prep Kit for Illumina. Sequencing was realized on an Illumina HiSeq 2500 System.
Project description:Analysis of Foxd3-regulation of self-renewal and quiescence in CDCP1+CSCs at gene expression level. The hypothesis tested in the present study was that the Foxd3 global developmental regulator serves a critical role to support the self-renewal of CDCP1+CSCs as one of few known factors involved in maintaining their quiescence. Results provide important information of the response of colorectal CDCP1+CSCs to depletion of Foxd3, such as genes involved in regulation of cell cycle, cell proliferation, and stem-like properties.
Project description:Our findings demonstrate that CDCP1 is a novel modulator of HER2 signalling, and a biomarker for the stratification of breast cancer patients with poor prognosis GEP analysis of human breast cancer cell lines SKBR3 overexpressing CDCP1 and control.
Project description:To elucidate functions of CDCP1, KDM3B and SDC1 molecules in HS5 human marrow stromal cells, global gene expression profiles were obtained and analyzed after knocking down the gene transcripts by using siRNA technology. Global gene expression profiles were obtained from HS5 marrow stromal cells after knocking down mRNA of CDCP1, KDM3B or SDC1 by using siRNA technology.
Project description:Circulating tumor cells (CTCs) and disseminated tumour cells with mesenchymal traits are difficult to detect by epithelial marker proteins. Particularly, triple negative breast cancers (TNBC) that are prone to therapy failure release a subpopulation of circulating tumour cells (CTCs) with mesenchymal traits. To provide tools that support their detection and analysis, the cell line BC-M1 established from disseminated tumour cells in the bone marrow of a breast cancer patient and a bone metastasis subline of MDA-MB-231 were analysed. Mass spectrometry analysis revealed high levels of CUB domain-containing protein 1 (CDCP1) in BC-M1. CDCP1 was found in other carcinoma cell lines (MDA-MB-231, MDA-MB-468) and other DTC cell lines (LC-M1, PC-E1) as well. Peripheral blood mononuclear cells were virtually negative for CDCP1 by Western Blot and immunofluorescent staining. Presence of CDCP1 in CTCs was confirmed by CellSearch. Here, CDCP1 positive CTCs were detected in eight of 30 analysed breast cancer patients. For the isolation of CTCs from the blood of breast cancer patients, we established a sandwich magnetic-activated cell sorting (MACS). The extracellular domain of CDCP1 served for cell catching and the cytoplasmic domain of CDCP1 for immunofluorescent detection of CDCP1 in CTCs. We showed that the MACS approach is suitable for the isolation of EpCam/keratin negative breast cancer cells from the blood and isolated CDCP1 positive CTCs from breast cancer patients by MACS. Hence, our approach is particularly suited for the detection and isolation of CTCs from TNBC when low EpCam or keratin levels limit the application of conventional approaches.
Project description:Panc1 (human pancreatic adenocarcinoma cells) cells were transfected with control siRNA (targeting firefly luciferase, siLuc) or siRNA targeting GLI1 (siGLI1, Pool of four siRNAs). 72h following transfection, RNA was prepared for array analysis.
Project description:To further develop gene expression approach, we have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to identify circ2082 expression effect in different subpopulations of glioblastoma cells (G cells). Cells were transfected with either control or antisense oligos targeting circ2082. KD of circ2082 affects gene expression patterns in cell subtype specific pattern
Project description:The Piwi–piRNA complex (Piwi–piRISC) in Drosophila ovarian somatic cells represses transposons transcriptionally to maintain genome integrity; however, the underlying mechanisms remain obscure. We performed mRNA-seq analysis from OSCs transfected with siRNAs against CG3893, the known piRNA pathway genes, Piwi, Maelstrom, HP1a and Armitage, and the control (EGFP), and PolII ChIP-seqanalysis from OSCs transfected with siRNAs against CG3893, Piwi, Mael and the control (EGFP). This result indicates that CG3893 is a novel factor for primary piRNA pathway in OSCs.