LIM kinase-2 targeting as a possible anti-metastasis therapy.
BACKGROUND
Metastatic properties of tumors involve movement of cancerous cells from one place to another and tissue invasion. Metastatic cells have altered cell adhesion and movement that can be examined by in vitro chemotaxis assays. The Rho/ROCK/LIM kinase pathway is one of the major signaling pathways involved in tumor metastasis. It is involved in the regulation of the actin cytoskeleton. Using the randomized ribozyme library, we initially found that metastatic human fibrosarcoma cells harboring ribozyme specific for ROCK lose their metastatic properties. In this study, we have determined the effect of ribozymes specific for LIM kinase-2 on metastatic and proliferative phenotypes of human fibrosarcoma cells.


METHODS
We attempted to target LIM kinase-2 (LIMK-2) expression by hammerhead ribozymes (Rz) in human metastatic fibrosarcoma cells. An effective ribozyme was selected based on the expression analysis. Cells were stably transfected with Rz specifically effective for LIMK-2 and were examined for metastatic and proliferative properties.


RESULTS
Analyses of cellular phenotypes such as cell proliferation, cell migration and colony-forming efficiency revealed that the suppression of LIMK-2 expression in human fibrosarcoma cells limits their migration and dense colony-forming efficiency without affecting cell proliferation rate or viability.


CONCLUSIONS
Specific targeting of metastatic and malignant properties of tumor cells by LIMK-2 ribozyme may serve as an effective therapy for invasive tumors with minimum effect on the surrounding normal cells.
Suyama
Eigo
E
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Tokyo 113-8656, Japan.
Wadhwa
Renu
R
Kawasaki
Hiroaki
H
Yaguchi
Tomoko
T
Kaul
Sunil C
SC
Nakajima
Motowo
M
Taira
Kazunari
K
D001483:Base Sequence; D045744:Cell Line, Tumor; D002465:Cell Movement; D002470:Cell Survival; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D054461:Lim Kinases; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009362:Neoplasm Metastasis / Q000628:therapy*; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases; D016337:RNA, Catalytic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D012366:RNA, Transfer, Val / Q000235:genetics; D014410:Tumor Stem Cell Assay
D004268:DNA-Binding Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012366:RNA, Transfer, Val; C427204:hammerhead ribozyme; C102895:LIMK2 protein, human; D054461:Lim Kinases; D017346:Protein Serine-Threonine Kinases