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. 2015 Jul 31;5:12460. doi: 10.1038/srep12460

Figure 4. miR-320a directly targets BCR/ABL.

Figure 4

(A) The 3′-UTR element of BCR/ABL messenger RNA was partially complementary to miR-320a. miR-320a, anti-miR-320a or scramble control and luciferase reporter containing either a wild type or a mutant 3′-UTR were co-transfected into HEK-293T cells. And a Renilla luciferase expressing construct exerts as internal control. (B) Western blot analysis of BCR/ABL expression in K562 cells infected with miR-320a, and NC transfected with miR-320a inhibitors (Anti-miR-320a). The gels have been run under the same experimental conditions. (C). Analysis of correlation of miR-320a and BCR/ABL expression in CML cancer stem cells and normal MSCs. *p < 0.05, **p < 0.01, ***p < 0.001. The analysis indicated that BCR/ABL and miR-320a were negatively correlated. (D–F) BCR/ABL abrogated the suppressive roles of miR-320a in K562 invasion and growth. K562 cells stably expressing miR-320a or scramble were transfect with or without BCR/ABL plasmids. Invasion assays (D), apoptosis analysis (E) and cell proliferation analysis (F) were performed with the above cells as described in Materials and Methods. Data are presented as mean ± s.e.m from at least three independent experiments. (G) Spearman’s correlation scatter plot of the levels of miR-320a (determined by in situ hybridization) and BCR/ABL protein (determined by immunohistochemistry) in 90 CML specimens. Representative images of BCR/ABL expression by immunohistochemistry were shown. Original magnification: ×200.