title: Gastric cancer cell proliferation is suppressed by frizzled-2 short hairpin RNA.
abstract: In order to identify novel targets for the molecular therapy of gastric cancer (GC), we investigated the mRNA and protein expression of frizzled-2 (Fz2), a Wnt signaling pathway receptor. Reverse-transcriptase polymerase chain reaction (PCR) amplification was utilized to determine the expression patterns of Fz genes in normal stomach and in the GC cell lines MKN45 and MKN74. Immunostaining was performed on surgical specimens of GC using an antibody against Fz2. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay was performed on MKN45 cells and MKN74 cells transfected with Fz2 short-hairpin (sh) RNA. Cell motility was analyzed by scratch assay following Fz2 shRNA. Real-time quantitative PCR was performed to analyze the expression levels of cyclin D1 and matrix metallopeptidase 9 (MMP-9). Fz1, 3, 6 and 8 were expressed in normal stomach, and in MKN45 and MKN74 cells. Fz2 was expressed in normal stomach and in MKN45, but not in MKN74 cells. Well-differentiated GC tissue was weakly positive for Fz2 in cell membranes. Fz2 was positive in both the cell membrane and cytoplasm of GC tissues of moderately differentiated and poorly differentiated adenocarcinoma. Signet ring cells were positive for cytoplasmic Fz2. Proliferation of MKN45 and MKN74 cells was suppressed by Fz2 shRNA, and a scratch assay demonstrated that Fz2 shRNA suppressed also MKN45 and MKN74 cell motility. Furthermore, Fz2 shRNA application led to downregulated mRNA expression of both cyclin D1 and MMP-9. Fz2, 3, 6 and 8 were expressed in normal stomach, and in MKN45 and MKN74 GC cells. Fz2 shRNA suppressed cell proliferation and motility of MKN45 and MKN74 cells, and downregulated cyclin D1 and MMP-9 expression in these GC cell lines.
authors.affiliation: Department of Gastroenterology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Tomizawa
authors.affiliation: Department of Radiology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Fuminobu
authors.initials: F
authors.lastname: Shinozaki
authors.affiliation: Department of Neurology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Motoyoshi
authors.affiliation: Department of Rheumatology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Sugiyama
authors.affiliation: Department of Pediatrics, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Shigenori
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Neurosurgery, National Hospital Organization, Shimoshizu Hospital, Yotsukaido City, Chiba 284-0003, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Ishige
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000368:Aged; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D003593:Cytoplasm / Q000378:metabolism; D005260:Female; D051157:Frizzled Receptors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008875:Middle Aged; D034741:RNA, Small Interfering; D012016:Reference Values; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology*; D014162:Transfection
chemical_list: C489742:FZD2 protein, human; D051157:Frizzled Receptors; D034741:RNA, Small Interfering