Identification of metastasis-related genes in a mouse model using a library of randomized ribozymes.
Libraries of randomized ribozymes have considerable potential as tools for the identification of functional genes critically involved in a biological phenotype of interest in vitro. We have used a ribozyme library in an in vivo mouse model to identify genes related to metastasis. We injected weakly metastatic melanoma cells that had been treated with the library intravenously into mice. We then isolated ribozymes that accelerated metastasis from pulmonary tumors that had developed from metastasizing cells. As candidates for metastasis-related genes that were targets of the isolated ribozymes, we identified five unknown and three known genes: stromal interaction molecule 1 (STIM1), polymerase gamma2 accessory subunit (Polg2), and cytochrome P450, family 2, subfamily d, polypeptide 22 (Cyp2d22). Repression of four of these by small interfering RNAs indeed resulted in the accelerated mobility of cells in in vitro scratch-wound assay. The further characterization of these candidate genes would provide clues to the complex mechanism(s) of metastasis.
Suyama
Eigo
E
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Wadhwa
Renu
R
Kaur
Kamaljit
K
Miyagishi
Makoto
M
Kaul
Sunil C
SC
Kawasaki
Hiroaki
H
Taira
Kazunari
K
D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels; D045744:Cell Line, Tumor; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D000072467:Cytochrome P450 Family 2; D000074002:DNA Polymerase gamma; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D020868:Gene Silencing; D005821:Genetic Techniques; D005822:Genetic Vectors; D008175:Lung Neoplasms / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D009690:Nucleic Acid Conformation; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000737:chemistry; D016337:RNA, Catalytic / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000378:metabolism; D011897:Random Allocation; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071737:Stromal Interaction Molecule 1; D014162:Transfection; D014945:Wound Healing
D015220:Calcium Channels; D008562:Membrane Glycoproteins; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; C100757:Stim1 protein, mouse; D000071737:Stromal Interaction Molecule 1; D012313:RNA; D003577:Cytochrome P-450 Enzyme System; C414739:Cyp2d22 protein, mouse; D000072467:Cytochrome P450 Family 2; D000074002:DNA Polymerase gamma; D004259:DNA-Directed DNA Polymerase