WNT5A is a Key Regulator of the Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties in Human Gastric Carcinoma Cells Pathobiology 2013;80:235-244.

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WNT5A is a Key Regulator of the Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties in Human Gastric Carcinoma Cells Pathobiology 2013;80:235-244 - DOI:10.1159/000346843 Fig. 1. Expression of WNT5A and the effects of transduction of pWNT5A in human GC-derived cells. a Expression of WNT5A transcripts in GC cell lines (RT-PCR). The mRNA levels of GAPDH expression were analyzed as a loading control. b Expression of WNT5A protein in GC cell lines (Western blot). β-Actin was used as a control. c Representative illustrations of WNT5A expression in MKN-7 and TMK-1 cells. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). d Cell proliferation assay in MKN-7 cells after transfection of pWNT5A and EV. A representative illustration of the pWNT5A transfectant is shown in the upper left corner. e Cell motility assay. Left: representative illustration of migration of MKN-7 cells in the presence or absence of pWNT5A transfection. © 2013 S. Karger AG, Basel

WNT5A is a Key Regulator of the Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties in Human Gastric Carcinoma Cells Pathobiology 2013;80:235-244 - DOI:10.1159/000346843 Fig. 2. WNT5A induces EMT and increases CSC marker CD133 levels in MKN-7 cells. a Results of flow cytometric analysis. Percentages of each fraction (subG1, G1, S or G2/M) are shown in the upper right corner of each fluorogram. For the suppression of WNT signaling, WNT inhibitor XAV939 was added to the culturing media. b Results of real-time RT-PCR in MKN-7 cells in the presence or absence of pWNT5A transfection and/or treatment with XAV939. The mRNA levels of GAPDH expression were analyzed as a loading control. * p < 0.05; ** p < 0.01. EV = empty vector. c Expression of WNT5A, SNAI1 and CDH1 proteins in pWNT5A transfectant. β-Actin was used as a control. d Representative illustration of the tumorigenicity test. Note that subcutaneous tumor developed in the SCID mouse inoculated with pWNT5A-transfected MKN-7 cells. © 2013 S. Karger AG, Basel

WNT5A is a Key Regulator of the Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties in Human Gastric Carcinoma Cells Pathobiology 2013;80:235-244 - DOI:10.1159/000346843 Fig. 3. Knockdown of WNT5A suppresses gene expression related to the EMT and CSCs. a Knockdown of WNT5A was performed using a lentiviral shRNA transduction system. Altered levels of WNT5A and EMT-related proteins were detected by Western blot. b Expression of EMT- and CSC-related genes in shWNT5A-infected MKN-7 cells. The mRNA levels of GAPDH expression were analyzed as a loading control. c Results of a cDNA microarray analysis of gene expression involved in intracellular signaling, chemokine-cytokine interaction and focal adhesion. © 2013 S. Karger AG, Basel

WNT5A is a Key Regulator of the Epithelial-Mesenchymal Transition and Cancer Stem Cell Properties in Human Gastric Carcinoma Cells Pathobiology 2013;80:235-244 - DOI:10.1159/000346843 Fig. 4. Representative illustrations of immunohistochemistry of WNT5A in human GC tissues (intestinal- and diffuse-type GC cases). Note that high expression of WNT5A was detected in the peripheral site of cancer nests in intestinal-type GC. In diffuse-type GC, high WNT5A expression was detected homogenously. © 2013 S. Karger AG, Basel