Figure 2.
miR-885-5p inhibits neuroblastoma proliferation and survival. (a) TP53 wild-type IMR32, SH-EP, KELLY and TP53 mutant HDN33, SK-N-BE(2)c cells were transfected with miR-885-5p, miR-331-3p mimics, control miRNA (each 30 n), Lipofectamine 2000 (vehicle) or left untreated. Cell viability was measured by Alamar Blue assay. The data are represented as the means±S.D. (a; insert) SA-ß-Gal assays were performed 4 days after transfection; the positively stained cells were counted (mean percentages±S.D. of three experiments are presented). (b) IMR32, SH-EP, KELLY and SK-N-BE(2)c were transfected with miRNA mimics and assayed for anchorage-independent growth in soft agar after 2 weeks in culture. Colonies were fixed and stained with crystal violet and counted using ImageJ software. The graphs (right) show mean percentages of colonies number±S.D. of three experiments. (c) Cell cycle profiles of controls and cells transfected with miRNA mimics were determined using FACS analysis. (d) The sub-G1 fraction of neuroblastoma cell lines transfected with miRNA mimics or control miRNA transfectants were detected at 4-days post transfection using FACS analysis by detection of cells with a hypodiploid DNA content (bar=mean percentage of sub-G1±S.D.). Phosphatidylserine externalization was analyzed by annexin V/PI staining, the results show the percentage of annexin V-positive/PI negative and annexin V/PI double positive cells