Figure 1. Differential expression of miRNAs between MM cells treated with PRIMA-1^Met or DMSO control.

A. MM.S or 8226 cells were treated with PRIMA-1^Met (10 or 20 μM, respectively). After 8h cells were harvested to isolate total RNA including miRNA. miRNA was reverse transcribed followed by qPCR array analysis in a 96-well plate targeting the cancer pathway finder (MM.1S) or apoptosis pathway (8226). Data were analysed by the online software (SABiosciences) to see the differential expression of the miRNAs. B and C. cDNAs were further used to validate the expression of miRNAs (miRNA-29a, miRNA-29b, and miRNA-34a) in MM.1S (B) and 8226 (C) cells. Fold-changes of the genes are shown after normalizing the data with to a set of housekeeping genes. D and E. miRNA-29a expression in MM patient samples and normal hematopoietic cells. (D) miRNA-29a expression is significantly higher in normal peripheral blood mononuclear cells (PBMC) or bone marrow mononuclear cells (BMNC#1) compared to MM patient samples and cell lines. RNAs were isolated from purified patient MM cells, PBMCs, and BMNCs from healthy donors, and the cultured MM cell lines, followed by analysis of basal expression level of miRNA-29a using qRT-PCR. Raw Ct values were normalized to housekeeping SNORD61 and relative expression was calculated using the comparative Ct methods. Results shown are means ±SD. (E) MM.1S and 8226 cells were treated with PRIMA-1^Met (10 and 20 μM, respectively), MIRA-1 (10 and 20 μM, respectively), dexamethasone (5 μM), or doxorubicin (5 μM). Control cells were treated with DMSO. Eight hours after treatment, cells were harvested for RNA isolation and expression of miRNA by qPCR as described earlier.