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Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: RETRACTED ARTICLE: Exosomal microRNA-32-5p induces multidrug resistance in hepatocellular carcinoma via the PI3K/Akt pathway

Fig. 7

Exosomal miR-32-5p induces multidrug resistance in vitro and in vivo. a Schematic diagram of exosome extraction. b The sizes and numbers of exosomes were determined by a Delsa Nano Analyzer. c Western blots for the expression of the exosome markers Flotillin-1, TSG-101, and CD63. d miR-32-5p was overexpressed in exosomes from Bel/5-FU. e Real-time PCR for the expression of miR-32-5p in exosomes treated with ethanol and GW-4869. f Real-time PCR for the expression of miR-32-5p and PTEN in Bel7402 cells treated with PBS, exosomes from Bel/5-FU, mimics NC and mimics. g Exosomes from Bel/5-FU but not from Bel7402 induced multidrug resistance, and miR-32-5p inhibitor reversed resistance in Bel7402 cells. (h and k) Western blots for the expression of molecules of the PI3K/Akt pathway, E-cad and N-Cad in Bel7402 cells after treatment with PBS, exosomes from Bel7402 and Bel/5-FU; in Bel7402 cells into which miR-32-5p inhibitor or inhibitor NC was transferred after treatment with exosomes; and in Bel7402 cells transfected with miR-32-5p mimics and the respective NC. (i and j) Migration and invasion assay of Bel7402 cells after treatment with PBS, exosomes from Bel7402 and Bel/5-FU; Bel7402 cells into which miR-32-5p inhibitor or inhibitor NC was transferred after treatment with exosomes; and Bel7402 transfected with miR-32-5p mimics and the respective NC. (l) Growth curves of xenograft tumors derived from Bel7402 cells injected with exosomes and then treated with PBS or 5-FU. n = three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test. (m) IHC staining for PTEN, p-Akt, p-mTOR, p-P70S6K, Ki67, N-Cad, E-Cad, and CD31 in xenograft tumors. Original magnification, 400×; scale bar, 25 μm. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test. N-Cad, N-Cadherin; E-Cad, E-Cadherin; MVD, microvascular density. (n) Schematic diagram summarizing how exosomal miR-32-5p induces multidrug resistance via the PTEN/PI3K/Akt pathway through angiogenesis and EMT. Exo, exosome

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