Silencing of autocrine motility factor induces mesenchymal-to-epithelial transition and suppression of osteosarcoma pulmonary metastasis.
Phosphoglucose isomerase (PGI) is a multifunctional enzyme that functions in glucose metabolism as a glycolytic enzyme catalyzing an interconversion between glucose and fructose inside the cell, while it acts as cytokine outside the cell, with properties that include autocrine motility factor (AMF)-regulating tumor cell motility. Overexpression of AMF/PGI induces epithelial-to-mesenchymal transition with enhanced malignancy. Recent studies have revealed that silencing of AMF/PGI resulted in mesenchymal-to-epithelial transition (MET) of human lung fibrosarcoma cells and breast cancer cells with reduced malignancy. Here, we constructed a hammerhead ribozyme specific against GUC triplet at the position G390 in the human, mouse, and rat AMF/PGI mRNA sequence. Mesenchymal human osteosarcoma MG-63, HS-Os-1, and murine LM8 cells were stably transfected with the ribozyme specific for AMF/PGI. The stable transfectant cells showed effective downregulation of AMF/PGI expression and subsequent abrogation of AMF/PGI secretion, which resulted in morphologic change with reduced growth, motility, and invasion. Silencing of AMF/PGI induced MET, in which upregulation of E-cadherin and cytokeratins, as well as downregulation of vimentin, were noted. The MET guided by AMF/PGI gene silencing induced osteosarcoma MG-63 to terminally differentiate into mature osteoblasts. Furthermore, MET completely suppressed the tumor growth and pulmonary metastasis of LM8 cells in nude mice. Thus, acquisition of malignancy might be completed in part by upregulation of AMF/PGI, and waiver of malignancy might also be controlled by downregulation of AMF/PGI.
Niinaka
Yasufumi
Y
Department of Oral and Maxillofacial Surgery, Faculty of Medicine, University of Yamanashi, Chuo, Japan. yninaka@yamanashi.ac.jp
Harada
Kiyoshi
K
Fujimuro
Masahiro
M
Oda
Masamitsu
M
Haga
Arayo
A
Hosoki
Misa
M
Uzawa
Narikazu
N
Arai
Naoya
N
Yamaguchi
Satoshi
S
Yamashiro
Masashi
M
Raz
Avraham
A
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000556:secondary; D008969:Molecular Sequence Data; D012516:Osteosarcoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D047368:Tumor Burden; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
D015820:Cadherins; D016337:RNA, Catalytic; D014746:Vimentin; D007633:Keratins; D005956:Glucose-6-Phosphate Isomerase