Studying the effects of actin cytoskeletal destabilization on cell cycle by cofilin overexpression |
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Authors: | Yi-Jang Lee Peter C Keng |
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Institution: | (1) Department of Pathology and Laboratory Medicine, 9 Nothwood Ln, 06457 Middletown, CT;(2) Cancer Center, University of Rochester School of Medicine and Dentistry, 14642 Rochester, NY |
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Abstract: | The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However,
it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we
have established a drug-free system using cofilin overexpression to investigate the relationship between actin filaments and
cell cycle progression. Cofilin is a member of the actin depolymerization factor (ADF)/cofilin family, cofilin cDNA was cloned
to a tetracycline-inducible gene expression vector and stably transfected to human lung cancer H1299 epithelial cells. Destabilization
of actin filaments and morphological change was detected in cofilin overexpressing cells by actin analysis and microscopy,
respectively. Measurements of growth rates showed that cell proliferation was retarded in cells with overexpressed cofilin.
Also, cell cycle analysis showed that approx 90% of cofilin overexpressing cells were arrested in G1 phase, which is consistent
with previous reports that drug-mediated disruption of actin filaments can cause G1 phase arrest. Taken together, cofilin
overexpression cell model provides evidence that the effects of actin cytoskeletal destabilization on cell cycle progression
can be studied using molecular approach instead of drug. |
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Keywords: | Actin cytoskeleton tet-on gene expression system cofilin cell cycle G1 phase arrest |
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