Fig. 4 | Cell Death & Differentiation

Fig. 4

From: A p53/miR-30a/ZEB2 axis controls triple negative breast cancer aggressiveness

Fig. 4

miR-30a (5p/3p) overexpression results in a reduced BC cell migration in vitro and in a zebrafish xeno-transplantation model. a In vitro cell migration assay of MDA231 engineered to ectopically express miR-30a-5p, miR-30a-3p or control miRNA (miRCTR) in a ZEB2-proficient (siCTR) or ZEB2-deficient (si1-ZEB2 and si2-ZEB2) context (left panel). The right panel shows the migration capacity of MDA231 engineered to stably overexpress ZEB2 or GFP, used as a negative control. b In vitro cell migration assay of MDA231 transfected with anti-miR-30a-5p (A-miR-30a-5p), anti-miR-30a-5p (A-miR-30a-5p) or control anti-miRNA (A-CTR). The percentage of transmigrated cells was measured at 7 h post seeding. Data represent the mean of three independent experiments; *p < 0.05. c Scatter plot representing cell dissemination in zebrafish embryos of mCHERRY MDA231 transiently silenced for ZEB2 (si1-ZEB2 and si2-ZEB2) or engineered to stably express miR-30a (5p/3p). Cells transfected with an empty vector were used as a control (ctr). Cells were implanted in the yolk sac of 2-day-old embryos (fli1:EGFP strain). Embryos were automatically imaged at 6 dpi. Dots represent single cells; colors identify each microinjected embryo; x-axis indicates the migration from the injection point (0,0) toward the head (positive values) or the tail (negative values); n indicates the number of embryos analyzed. d Spreading distance of MDA231 and MDA157 calculated from data represented in (c) and in Supplementary Figure S8b, respectively; *p < 0.05. e Representative images of zebrafishes injected with MDA231 cells at 6 dpi. Cells were injected into the blood circulation (duct of Cuvier) of 2-day-old zebrafish embryos. Scale bar = 100 µm. f Percentages of embryos that show caudal micrometastatic colonization after injection with MDA231 or MDA157 cells engineered with control vector (ctr), miR-30a (5p/3p) or silenced for ZEB2 (si1-ZEB2 and si2-ZEB2). The percentage of ctr embryos showing metastasis was arbitrarily set to 100. Data are shown at 1, 4 and 6 dpi. Figures represent the results of two independent experiments

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