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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: The U2AF2 /circRNA ARF1/miR-342–3p/ISL2 feedback loop regulates angiogenesis in glioma stem cells

Fig. 3

The miR-342–3p negatively regulated ISL2 and miR-342–3p-mediated GCM suppressed the proliferation, invasion, and angiogenesis of hBMECs. a Identification of a miRNA that potentially regulated ISL2 expression based on microRNA, miRDB, Starbase, and TargetScan databases. b Schematic diagram of the putative miR-342–3p binding site in the 3′-UTR of ISL2. c The qPCR showed the mRNA expression correlation between ISL2 and miR-342–3p in 70 cases of glioma patients. d, g The luciferase reporter assays showed that miR-342–3p mimic (d) or inhibitor (g) altered the luciferase promoter activities of ISL2. e, f Western blotting (e) and qPCR (f) showed the expression of ISL2 in glioma stem-like cells (GSCs) after miR-342–3p mimic or inhibitor treatment. h MTS assays showed that the miR-342–3p mimic or inhibitor transfected GCM affected hBMEC viability and was reversed by ISL2 overexpression or knockdown, respectively. i The EDU assay showed that the miR-342–3p mimic or inhibitor transfected GCM affected the proliferation of hBMECs and was reversed by ISL2 overexpression or knockdown, respectively. Scale bar = 50 μm. g A representative Transwell assay showed that the miR-342–3p mimic or inhibitor-transfected GCM affected the invasion of hBMECs and was reversed by ISL2 overexpression or knockdown, respectively. Scale bar = 100 μm. k Representative tube formation assay showed that the miR-342–3p mimic or inhibitor-transfected GCM affected the tubulogenesis of hBMECs and was reversed by ISL2 overexpression or knockdown, respectively. Scale bar = 100 μm. l, m The qPCR (l) and ELISA assay (m) indicated that the miR-342–3p mimic or inhibitor regulated mRNA expression and secretion of VEGFA in GSCs and was reversed by ISL2 overexpression or knockdown, respectively. EV: empty vector, OE: overexpression, NC: negative control, KD: knockdown. All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001

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