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Identification of genes responsible for cell migration by a library of randomized ribozymes
Eigo Suyama1, Hiroaki Kawasaki, Tatsuhiko Kasaoka
1Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Tokyo 113-8656, Japan.
Abstract:
Several genes appear to be associated with metastasis, but the underlying mechanisms of metastasis still remain unclear. In this study, we used a library of randomized ribozymes to identify, by inactivation of transcripts, genes involved in cell migration that is an essential aspect of metastasis. Using a chemotaxis assay, the ribozymes that inhibited cell migration were selected from the library. Among such ribozymes, we found two ribozymes that targeted and cleaved ROCK1 mRNA at independent sites. ROCK1 and ROCK2 are Rho kinases, and it has been demonstrated that they regulate the organization of the actin cytoskeleton and are responsible for cell motility and cytokinesis. The two ribozymes that specifically cleaved ROCK1 mRNA inhibited both the migration and invasion of invasive HT1080 fibrosarcoma, but neither had any effect on cell proliferation. Our analysis indicates that the ribozymes toward ROCK1 can block invasive activity but not the proliferation of HT1080 cells without having any effect on expression of ROCK2. Ribozymes identified in this study, including the ribozymes against ROCK1, might be useful in understanding the mechanisms of cell migration and metastasis.
Insights
Researchers identified ribozymes that target ROCK1 mRNA, effectively inhibiting cancer cell migration and invasion without affecting proliferation. This discovery offers new insights into metastasis mechanisms.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Background:
- Metastasis mechanisms remain unclear despite identified associated genes.
- Cell migration is a critical component of metastasis.
Purpose of the Study:
- To identify genes involved in cell migration using a ribozyme library.
- To investigate the role of Rho kinases in metastasis.
Main Methods:
- Utilized a randomized ribozyme library for transcript inactivation.
- Employed a chemotaxis assay to select ribozymes inhibiting cell migration.
- Identified ribozymes targeting and cleaving ROCK1 mRNA.
Main Results:
- Two ribozymes specifically cleaved ROCK1 mRNA at independent sites.
- These ribozymes inhibited migration and invasion of HT1080 fibrosarcoma cells.
- Neither ribozyme affected cancer cell proliferation or ROCK2 expression.
Conclusions:
- Ribozymes targeting ROCK1 effectively block invasive activity without impacting proliferation.
- Identified ribozymes provide tools for understanding cell migration and metastasis mechanisms.