Project description:Forced expression of Bmi1 accelerated the self-renewal of hepatic stem/progenitor cells and eventually induced their transformation in an in vivo transplant model. The Ink4a/Arf locus, which encodes a cyclin-dependent kinase inhibitor, p16Ink4a, and a tumor suppressor, p19Arf, is a pivotal target of Bmi1. Therefore, it would be of importance to understand the contribution of the Ink4a/Arf locus to Bmi1 oncogenic functions in cancer and search for as-yet-unknown Bmi1 target genes other than Ink4a/Arf. We used microarrays to explore novel candidate downstream targets for Bmi1 in hepatic stem/progenitor cells Experiment Overall Design: Purified Dlk-positive hepatoblasts at day 28 of culture were subjected to RNA extraction and hybridization on Affymetrix microarrays. Data were obtained for quadrant samples from four independent experiments.
Project description:BMI1-silenced Prostate cancer cells exhibited a significant change in the expression of several proliferation-associated genes. These included Cyclin-D1, BCL2, IL and NF-kappaB. An increased expression of p16, p15 and TIMP3 in BMI1-silenced tumor cells was observed List of Selected Genes Modulated by BMI1-supression in Prostate cancer Cells qPCR gene expression profiling. LNCaP and BMI1-silenced LNCaP cells were used as indicated in the summary. Equal amount of total RNA from each group was pooled prior to gene expression analysis.
Project description:Neural stem cells were isolated from adult mouse subventricular zone and transduced with a Bmi1-overexpressing lentiviral vector or an empty vector control. Cells were grown as neurospheres (in non-adherent culture conditions) for three passages and RNA purifed (after four weeks).
Project description:The polycomb group proteins form large repressive complexes. It has recently become evident, via ChIP on chip analysis, that the polycomb complexes, PRC1 and PRC2, target a large number of genes (Boyer et al., 2006; Bracken et al., 2006; Tolhuis et al., 2006), which are involved in cell fate decisions. In order to identify genes which are truly regulated by the polycomb group proteins, CBX8 and BMI1, we expressed these two proteins in mouse embryonic fibroblasts (MEFs) by retroviral infection.
Project description:Bmi1 is a component of the Polycomb-repressive complexes (PRC) and essential for maintaining the pool of adult stem cells. PRC are key regulators for embryonic development by modifying chromatin architecture and maintaining gene repression. To assess the role of Bmi1 in pluripotent stem cells and upon exit from pluripotency during differentiation, we studied forced Bmi1 expression in mouse embryonic stem cells (ESC). We found that ESC do not express detectable levels of Bmi1 RNA and protein and that forced Bmi1 expression had no obvious influence on ESC self-renewal. However, upon ESC differentiation Bmi1 effectively enhanced development of hematopoietic cells. Global transcriptional profiling identified a large array of genes that were differentially regulated during ESC differentiation by Bmi1. Importantly, we found that Bmi1 induced a prominent up-regulation of Gata2, a zinc finger transcription factor, which is essential for primitive hematopoietic cell generation from mesoderm. In addition, Bmi1 caused sustained growth and a more than 100-fold expansion of ESC-derived hematopoietic stem/progenitor cells within 2-3 weeks of culture. The enhanced proliferative capacity was associated with reduced Ink4a/Arf expression in Bmi1-transduced cells. Taken together, our experiments demonstrate distinct activities of Bmi1 in ESC and ESC-derived hematopoietic progenitor cells. In addition, Bmi1 enhances the propensity of ESC in differentiating towards the hematopoietic lineage. Thus, Bmi1 could be a candidate gene for engineered adult stem cell derivation from ESC. 8 samples in total. Bmi1 embryonic stem cells sample_1 (Bmi1_ESC_1) Bmi1 embryonic stem cells sample_2 (Bmi1_ESC_2) Untreated CCE embryonic stem cells (CCE_ESC_Control) Empty vector CCE embryonic stem cells (CCE_ESC_Vector) Bmi1 embryoid body sample_1 (Bmi1_EB_1) Bmi1 embryoid body sample_2 (Bmi1_EB_2) Empty vector control embryoid body sample_1 (Vector_EB_1) Empty vector control embryoid body sample_2 (Vector_EB_2)
Project description:Neural stem/progenitor cells were isolated from the lateral ventricle wall of 4-6 week-old CD1 mice and grown as neurospheres under low density culture conditions. Test cells were transduced with bicistronic retroviral constructs for the over-expression of Bmi1 together with eGFP, and control cells were transduced with an empty vector construct expressing eGFP only. To identify genes, which are regulated by BMI1 in neural stem/progenitor cells, the gene expression profiles of neurosphere cells over-expressing Bmi1 were compared empty vector control cells using Affymetrix Gene mouse ST1.0 arrays 3 independent Bmi1-overexpressing neurosphere cultures (test samples) were compared to 3 independent empty vector neurosphere cultures (control samples).
Project description:The purpose of our study was to examine the function of BMI1 in fusion positive and fusion negative rhabdomyosarcoma cell lines and validate these findings in patient tissue. RD and RH30 cell lines were treated with control-siRNA or BMI1-siRNA and the RNA was subsequently sequenced on a HiSeq2500. The resultant sequencing data was used for Gene Set Enrichment Analysis and pathway analysis.
Project description:Transcriptional profiling of Bmi1 mutant dental epithelia including the stem cell compartment to determine which genes are upregulated in response to loss of Bmi1. Two condition experiment: dental epithelia homozygous null for Bmi1 and WT dental epithelia. 4 replicates each