Expression profiling of SMMC-7721 cells: stably expressing lncRNA-hPVT1 vs LV-Control
ABSTRACT: In our present study, we found that lncRNA-hPVT1 could promote HCC cell proliferation. We want to know what were the target genes of lncRNA-hPVT1. So we constructed the hPVT1-overexpressed SMMC-771 cells and observed the mRNA profile in hPVT1-overexpressed and control SMMC-771 cells. lncRNA-hPVT1 induced transcriptional changes in SMMC-7721. Two-condition experiment, control (LV-Control) vs lncRNA-hPVT1 expression (LV-hPVT1 Clone2 ). Each has 3 biological repeats.
Project description:lncRNA-PRAL induced transcriptional changes in SMMC-7721 Overall design: Two-condition experiment, control (AV-CON) vs lncRNA-PRAL expression (AV-PRAL). Each has 3 biological repeats.
Project description:To further development of the effects of miR-200a in ovarian cancer OVCAR3 cells，we have employed lncRNA and mRNA microarray as a discovery platform to identify lncRNA and mRNA expression in miR-200a overexpressing ovarian cancer cells. OVCAR3 cells were transfected with lentiviral vector with eGFP, encoding miR-200a and negative control vector (LV- miR-200a and LV-CON,) by using polybrene. The dysregulation of miR-200a was confirmed by using RT-PCR. RNA was extracted and detected by a lncRNA and mRNA microarray in LV-miR-200a and LV-CON OVCAR3 cells. The different expression of lncRNA and mRNA in LV-miR-200a and LV-CON OVCAR3 cells was analyzed to explore the mechanism that miR-200a affect ovarain cancer cells. Overall design: miR-200a overexpression induced lncRNA and mRNA expression in OVCAR3 cells was measured in LV-miR-200a and LV-CON groups. 3 control replicates at each group.
Project description:To further development of the effects of β-HCG in ovarian cancer SKOV3 cells，we have employed lncRNA and mRNA microarray as a discovery platform to identify lncRNA and mRNA expression in β-HCG overexpressing ovarian cancer cells. SKOV3 cells were transfected with lentiviral vector with eGFP, encoding β-HCG and negative control vector (LV- β-HCG and LV-CON,) by using polybrene. The dysregulation of β-HCG was confirmed by using RT-PCR. RNA was extracted and detected by a lncRNA and mRNA microarray in LV-β-HCG and LV-CON SKOV3 cells. The different expression of lncRNA and mRNA in LV-β-HCG and LV-CON SKOV3 cells was analyzed to explore the mechanism that β-HCG affect ovarain cancer cells. Overall design: β-HCG overexpression induced lncRNA and mRNA expression in SKOV3 cells was measured in β-HCG and LV-CON groups. 3 control replicates at each group.
Project description:Adult mouse subventricular zone (SVZ) neural progenitor cells (NPCs) retain the capacity to generate multiple lineages in vitro and in vivo. Thus far, the mechanisms involved in the regulation of these cells have not been well elucidated. We have carried out RNA profiling of adult SVZ cell cultures undergoing differentiation, to identify pathways that regulate progenitor cell proliferation and to define a set of transcripts that can be used as molecular tools in the drug discovery process. We carried out a stepwise stratification of the results to identify transcripts specifically enriched in NPCs and validated some of these using comparative literature analysis, quantitative PCR and immunological techniques. The results show a set of transcription factors, secreted molecules, and plasma membrane markers which are differentially regulated during differentiation. Pathway analysis highlights alterations in IGF, Wnt and TGFbeta signalling cascades. Further characterization of these components could provide greater insight into the mechanisms involved in the regulation of neurogenesis in the adult brain. Mouse subventricular zone neural stem cells cultures were prepared from C57Bl6/J mice (Johansson et al.,1999) and cultured in suspension in DFEM/F12 1:1 supplemented with PSF, B27 supplement and EGF/ FGF-2 at 20ng/ml. RNA was isolated from either expanding neurospheres in the presence of FGF2 and EGF (Reynolds and Weiss, 1992; Gritti et al., 1999), or from cultures induced to differentiate upon growth factor withdrawal or serum exposure, either after 24 hours (in suspension, to avoid changes induced by substrate attachment) or 5 days (as adherent cultures in poly-lysine and laminin-coated plates). Experiments were conducted in triplicate from independent batches of SVZ preparations. As a control for potential effects of media changes, a set of identical cultures were included where cells were grown for a further 24 hours in fresh media containing FGF2 and EGF (20 ng/ml each, normal growth media, NGM). Fluorescently labelled aRNA from individual samples were competitively hybridised against a pool of labelled aRNA from undifferentiated reference (starting) material on custom Agilent microarrays representing ~22,500 mouse transcripts. Technical, fluor-reversed hybridisations were performed for every sample.
Project description:Lung cancer cell line, A549 cells, was transfected with siPML or siCtrl for 48hr to knockdown PML expression. WDR4 or vector was overexpressed in A549 cells for 48hr. These four sets of cells were then subjected to microarray profiling using the Human OneArray microarray (version HOA6.1, GEO Platform GPL19137) from Phalanx Biotech Group. Comparison of transcriptomes from siPML/siCtrl and W4/vec in A549 cells
Project description:To explore more details in the similarities and differences between ABN+Rarg+Nr5a2 iN cells and primary neurons, we compared the global gene expression pattern of matured iN cells 12 days post infection with mouse primary neurons and MEFs by microarray analysis. In the study presented here, a total of 26423 unique genes were detected in primary mouse embryonic fibroblasts, induced neurons from MEFs by Ascl1+Brn2+Ngn2+Rarg+Nr5a2 at 12 days after infection and primary neurons from postnatal day 0 hippocampus, leading to more details about the similarities and differences among the three groups.