Fig. 1

Identification of miR-141 as a secretary miRNA from ovarian cancers. (A) The heatmap indicated the changes in secretary miRNA expression in ovarian cancer patients and normal donors using a miRCURY LNA miRNA PCR Array. N = 2 independent experiments (left). The graph showed the expression of secretory miRNAs compared between ovarian cancer patients and normal donors in the miRCURY LNA miRNA Cancer Focus PCR Panel (right). N = 2 independent experiments. (B) The graphical chart showed the elevated level of exosomal miR-141 in the serum of OvCa patients (n = 62) compared with normal women (n = 24) by qPCR analysis (mean ± SEM, t-test). (C) The graphical chart showed the secretory miR-141 in a panel of ovarian cancer cell lines by qPCR analysis. N = 1 independent experiment. (D) Knockdown of n-SMase2 by a lentiviral shRNAi approach led to the complete suppression of miR-141 exosome production in the conditioned medium of CAOV3 and OVCA433 cells. β-actin and U6 were the internal controls. (n = 3, mean ± SEM, t-test, **P < 0.01) (E) The immunofluorescence microscope showed PHK67 labeled exosomal miR-141 derived from OVCA433 conditioned medium, localized in the cytoplasm of WI-38 and T HESC stromal cells. Representative images are shown in color. Blue, DAPI staining in the nucleus. Green, staining of the exosomes with PKH67. Scale bar = 20 μm