Gene expression signatures for human interleukin-6 (IL-6) treatment in human multiple myeloma (MM) cell lines.
Ontology highlight
ABSTRACT: To investigate the effect of IL-6 on gene expression in IL-6-dependent MM cell lines, we have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to be associated with the cell proliferation of MM cells. ANBL-6 and FLAM-76 cells were incubated for 48 h in the absense of IL-6, and then the cells were treated with IL-6 (10 ng/mL) for 0, 6, 12, and 24 h in vitro.
Project description:To examine the effect of C1orf112 disruption on the gene expression changes in MM cells , we generated C1orf112-KO cells with KMS-11, OPM-2, and ANBL-6 cells. The control and C1orf112-KO MM cells were incubated for 48 h in the typical cell culture medium.
Project description:To investigate the relationship between the LEN resistance and gene expression profile, we generated LEN-resistant (LEN-R) cell clones using U266 and NCU-MM1 cell lines by chronically exposing low dose of LEN (0.1 to 10 μM). The parental and LEN-R MM cells were incubated with either DMSO or LEN (10 μM) for 24 h.
Project description:All-trans retinoic acid (ATRA) is important for sensitizing MM cells to carfilzomib (Cfz). To determine what signalling pathways are affected by ATRA in Cfz-treated MM cells, MM.1S MM cell line was pulsed with Cfz and then cultured with DMSO or 10µM ATRA for 12 h. Total RNAs of 2 x 106 MM.1S cells were extracted by RNeasy Mini Kit (Qiagen). 5-10 µg RNA samples were sent to Gene Expression and Genotyping Facility at Case Western Reserve University (Cleveland, OH) for genearray followed by data analysis. We use microarray data to determine differential expression of genes in Cfz-treated MM cells in culture with DMSO and ATRA.
Project description:Macophage migration inhibitor (MIF) is important for MM resistance to proteasome inhibitors. To determine what signalling pathways are affected by MIF in MM cells, we established MIF-KO MM.1S and ARP-1 MM cell lines. Total RNAs of 2 x 106 CTR-KO and MIF-KO MM cells were extracted by RNeasy Mini Kit (Qiagen). 5-10 µg RNA samples were sent to Gene Expression and Genotyping Facility at Case Western Reserve University (Cleveland, OH) for RNA-seq followed by data analysis. We use microarray data to determine differential expression of genes in MIF-KO and CTR-KO MM cells.
Project description:We have measured the proteomic changes on ANBL6, IH1, INA6, U266, KJON, OH2, VOLIN, LR5, KARPAS, RAMOS cell lines and used it to build a super SILAC library in order to quantify proteomic changes from MGUG to MM stages of Myeloma with Elite orbitrap mass spetrometer.
Project description:Recent studies have delineated cancer type-specific roles of histone 3 lysine 27 (H3K27) demethylase KDM6B/JMJD3 depending on its H3K27 demethylase activity. Here we show that KDM6B is expressed in multiple myeloma (MM); and that shRNA-mediated knockdown and CRISPR-mediated knockout of KDM6B abrogate MM cell growth and survival. TNFα or bone marrow stromal cell culture supernatants induce KDM6B, which is blocked by IKKβ inhibitor MLN120B, suggesting KDM6B is regulated by NF-κB signaling in MM cells. RNA-sequencing and subsequent ChIP-qPCR analyses reveal that KDM6B is recruited to the loci of genes encoding components of MAPK signaling pathway including ELK1 and FOS, and upregulates these genes expression without affecting H3K27 methylation level. Overexpression of catalytically-inactive KDM6B activates expression of MAPK pathway-related genes, confirming its function independent of demethylase activity. We further demonstrate that downstream targets of KDM6B, ELK1 and FOS, confer MM cell growth. Our study therefore delineates KDM6B function that links NF-κB and MAPK signaling pathway mediating MM cell growth and survival, and validates KDM6B as a novel therapeutic target in MM.
Project description:Recent studies have highlighted the oncogenic roles of KDM5A; however, its molecular mechanism has not been fully delineated. Here we identifies KDM5A as a transcriptional activator of MYC target genes in multiple myeloma (MM). Genetic ablation of KDM5A or pharmacologic inhibition of KDM5 by novel inhibitor JQKD82 impairs MM cell growth. Gene expression profile reveals that JQKD82 decreases expression of MYC targets. Expression of MYC targets is similarly downregulated by KDM5A depletion, but little affected by KDM5B knockdown. Mechanistically, KDM5A co-localizes with MYC across the genome, and KDM5A and MYC coordinately promote MYC target gene transcription. The treatment with JQKD82 reduces phosphorylation level of RNA polymerase II (RNAPII), resulting in RNAPII pausing, accompanied by hyper-H3K4me3 status at the MYC loci. Finally, we shows that JQKD82 exerts anti-MM activity and prolongs overall survival in vivo mouse xenograft models. Our results delineate KDM5A function that reinforces MYC transcriptional program, and identify KDM5A as a potential therapeutic target in MM.