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  • Comparison of the aneugenic properties of nocodazole, paclitaxel and griseofulvin in vitro. Centrosome defects and alterations in protein expression profiles.

Comparison of the aneugenic properties of nocodazole, paclitaxel and griseofulvin in vitro. Centrosome defects and alterations in protein expression profiles.

Journal of applied toxicology : JAT (2012-03-21)
Polyxeni Zacharaki, Georgia Stephanou, Nikos A Demopoulos
ABSTRACT

We have comparatively investigated the aneugenic activity of two anticancer drugs, nocodazole (NOC) and paclitaxel (PTX), and the antifungal griseofulvin with promising role in cancer treatment (GF), which affect microtubule dynamics in different ways. The comparison was achieved in HFFF2 human fibroblasts, MCF-7 human breast cancer cells and C2C12 mouse myoblasts, and focused on three issues: (i) induction of chromosome delay by estimation of MN frequency using CREST analysis; (ii) disturbance of spindle organization with Aurora-A/β-tubulin immunofluorescence; and (iii) alterations in the expression of Aurora-A, β- and γ-tubulin by western blotting. They induced chromosome delay, provoked metaphase arrest and promoted microtubule disorganization, reflecting their common characteristic of generating aneuploidy. In particular, NOC induced mainly monopolar metaphases, although PTX induced only multipolar metaphases. GF generated different types of abnormal metaphases, exhibiting cell specificity. Additionally, NOC decreased the expression of Aurora-A and β-tubulin, while the opposite held true for PTX and GF. γ-Tubulin expression was not modulated owing to NOC treatment, whereas PTX and GF increased γ-tubulin expression. Our findings throw a light on the manifestation of the aneugenicity of the studied compounds through centrosome proliferation/separation and protein expression, reflecting their different effects on microtubule dynamics.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Griseofulvin, from Penicillium griseofulvum, 97.0-102.0%
Sigma-Aldrich
Nocodazole, ≥99% (TLC), powder
Sigma-Aldrich
Anti-Mouse IgG (Fab specific)–TRITC antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Goat serum
Sigma-Aldrich
Monoclonal Anti-β-Tubulin antibody produced in mouse, clone TUB 2.1, ascites fluid
Sigma-Aldrich
Anti-Human Polyvalent Immunoglobulins (whole molecule)−FITC antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
Anti-Mouse IgG (whole molecule) F(ab′)2 fragment–FITC antibody produced in sheep, affinity isolated antibody, buffered aqueous solution