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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

PLAMED-2010-06-0597-OP.R2

Supporting Information

Protoflavonoids from Ferns Impair Centrosomal Integrity of Tumor Cells

Isabelle Pouny1, Chantal Etiévant2, Laurence Marcourt1, Isabelle Huc-Dumas1, Muriel

Batut1, Frédéric Girard2, Michel Wright2, and Georges Massiot1

Affiliation 1 Unité Mixte de Service 2597, Centre National de la Recherche Scientifique (CNRS) - Pierre

Fabre, 2 Unité Mixte de Recherche 2587, Centre National de la Recherche Scientifique (CNRS) - Pierre

Fabre,

Institut des Sciences et Technologies du Médicament de Toulouse (ISTMT), Toulouse, France

Correspondence

Dr. G. Massiot

Unité Mixte de Service 2597

Centre National de la Recherche Scientifique (CNRS) - Pierre Fabre, ISTMT

3 rue des Satellites

BP94244

31432 Toulouse Cedex 4

France

Tel. : +33/5/34321402

Fax: +33/5/34321414

georges.massiot@cnrs-dir.fr

Dr. M. Wright

Unité Mixte de Recherche 2587

Centre National de la Recherche Scientifique (CNRS) - Pierre Fabre

ISTMT, 3 rue des Satellites

BP94244

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

31432 Toulouse Cedex 4

France

Tel. : +33/5/34321364

Fax: +33/5/34321350

michel.wright@istmt.cnrs.fr

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Fig 1S 1H and 13C NMR spectra of compound 1 (CD3OD + CDCl3, 500MHz).

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Fig 2S 1H and 13C NMR spectrum of compound 2 (CD3OD, 500MHz).

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Table 1S List of ferns tested Plant name Family Plant part Mitose effect Psilotatae Psilotum triquetrum Psilotaceae Whole plant Lycopodiatae Huperzia gnidioides Lycopodiaceae Whole plant + Lycopodiella cernua Lycopodiaceae Whole plant Lycopodium annotinum Lycopodiaceae Aerial parts Lycopodium casuarinoides Lycopodiaceae Whole plant + Lycopodium cernuum Lycopodiaceae Aerial parts Lycopodium clavatum Lycopodiaceae Whole plant Lycopodium complanatum Lycopodiaceae Whole plant Lycopodium serratum Lycopodiaceae Whole plant Selaginella braunii Selaginellaceae Whole plant Selaginella doederleinii Selaginellaceae Whole plant Selaginella fissidentoides Selaginellaceae Whole plant Selaginella involvens Selaginellaceae Whole plant Selaginella kraussiana Selaginellaceae Whole plant Selaginella tamariscina Selaginellaceae Whole plant + Selaginella trichoclada Selaginellaceae Whole plant Selaginella uncinata Selaginellaceae Whole plant Equisetatae Equisetum arvense Equisetaceae Whole plant Equisetum arvense Equisetaceae Fresh aerial parts Equisetum debile Equisetaceae Aerial parts

Equisetum debile Equisetaceae Underground parts

Equisetum debile Equisetaceae Whole plant + Equisetum fluviatile Equisetaceae Stems + Equisetum hyemale Equisetaceae Whole plant Equisetum hyemale Equisetaceae Aerial parts

Equisetum hyemale Equisetaceae Underground parts

Equisetum palustre Equisetaceae Fresh aerial parts Equisetum pratense Equisetaceae Whole plant Equisetum ramosissimum Equisetaceae Whole plant + Equisetum ramosissimum Equisetaceae Fresh aerial parts Equisetum telmateia Equisetaceae Fresh aerial parts Filicatae Asplenium lineatum Aspleniaceae Leaves Asplenium nidus Aspleniaceae Leaves Asplenium prolongatum Aspleniaceae Whole plant Asplenium scolopendrium Aspleniaceae Aerial parts

Asplenium scolopendrium Aspleniaceae Underground parts

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Asplenium trichomanes* Aspleniaceae Aerial parts Asplenium trichomanes* Aspleniaceae Aerial parts Blechnum attenuatum Blechnaceae Leaves Blechnum attenuatum Blechnaceae Stems Blechnum montbrisonis Blechnaceae Leaves Blechnum montbrisonis Blechnaceae Stems Blechnum occidentale Blechnaceae Whole plant

Blechnum orientale Blechnaceae Underground parts

Blechnum orientale* Blechnaceae Aerial parts Blechnum orientale* Blechnaceae Aerial parts Blechnum spicant Blechnaceae Aerial parts Blechnum spicant Blechnaceae Whole plant Struthiopteris eburnea Blechnaceae Underground

parts + Struthiopteris eburnea Blechnaceae Leaves Woodwardia japonica Blechnaceae Leaves Woodwardia japonica Blechnaceae Stems

Woodwardia japonica Blechnaceae Underground parts

Woodwardia unigemmata Blechnaceae Underground parts

Cyathea borbonica Cyatheaceae Leaves Cyathea excelsa Cyatheaceae Leaves Cyathea pubescens Cyatheaceae Fronds Cyathea spectabilis Cyatheaceae Aerial parts Cyathea spinulosa Cyatheaceae Leaves Cyathea spinulosa Cyatheaceae Stems Davallia tyermanii Davalliaceae Aerial parts

Davallia tyermanii Davalliaceae Underground parts +

Dennstaedtia davallioides Dennstaedtiaceae Whole plant Dennstaedtia scabra Dennstaedtiaceae Whole plant Histiopteris incisa Dennstaedtiaceae Whole plant Microlepia marginata* Dennstaedtiaceae Whole plant Microlepia marginata* Dennstaedtiaceae Whole plant

Microlepia strigosa Dennstaedtiaceae Underground parts

Microlepia strigosa Dennstaedtiaceae Aerial parts Odontosoria chusana* Dennstaedtiaceae Whole plant + Odontosoria chusana* Dennstaedtiaceae Whole plant + Onychium japonicum Dennstaedtiaceae Whole plant Pteridium aquilinum Dennstaedtiaceae Leaves Pteridium aquilinum Dennstaedtiaceae Stems

Pteridium aquilinum Dennstaedtiaceae Underground parts

Pteridium revolutum Dennstaedtiaceae Leaves Pteridium revolutum Dennstaedtiaceae Underground

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

parts

Pteridium revolutum Dennstaedtiaceae Stems Cibotium barometz* Dicksoniaceae Leaves Cibotium barometz* Dicksoniaceae Leaves

Cibotium barometz Dicksoniaceae Underground parts

Cibotium barometz* Dicksoniaceae Stems Cibotium barometz* Dicksoniaceae Stems + Phymatodes scolopendria Dipteridaceae Whole plant Allantodia metteniana Dryopteridaceae Leaves Allantodia metteniana Dryopteridaceae Underground

parts Arachniodes chinensis Dryopteridaceae Whole plant Arachniodes sporadosora Dryopteridaceae Whole plant Athyrium filix-femina Dryopteridaceae Aerial parts Athyrium niponicum var pictum Dryopteridaceae Aerial parts

Athyrium niponicum var pictum Dryopteridaceae Underground parts

Deparia lobato-crenata Dryopteridaceae Underground parts

Deparia lobato-crenata Dryopteridaceae Aerial parts Diplazium esculentum Dryopteridaceae Stems Diplazium esculentum Dryopteridaceae Leaves Diplazium donianum Dryopteridaceae Whole plant

Diplazium subsinuatum Dryopteridaceae Underground parts

Diplazium subsinuatum Dryopteridaceae Aerial parts Dryopteris affinis Dryopteridaceae Fronds

Dryopteris affinis Dryopteridaceae Underground parts

Dryopteris carthusiana Dryopteridaceae Whole plant +

Dryopteris crassirhizoma Dryopteridaceae Underground parts

Dryopteris erythrosora Dryopteridaceae Aerial parts +

Dryopteris erythrosora Dryopteridaceae Underground parts

Dryopteris filix-mas* Dryopteridaceae Underground parts

Dryopteris filix-mas Dryopteridaceae Aerial parts

Dryopteris filix-mas* Dryopteridaceae Underground parts

Dryopteris laeta Dryopteridaceae Underground parts

Dryopteris laeta Dryopteridaceae Leaves Dryopteris sieboldii Dryopteridaceae Whole plant +

Dryopteris varia Dryopteridaceae Underground parts +

Dryopteris varia Dryopteridaceae Aerial parts + Dryopteris wallichiana Dryopteridaceae Underground +

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

parts

Matteuccia intermedia Dryopteridaceae Underground parts +

Matteuccia intermedia Dryopteridaceae Stems + Matteuccia intermedia Dryopteridaceae Leaves + Matteuccia orientalis Dryopteridaceae Whole plant + Matteuccia struthiopteris Dryopteridaceae Aerial parts

Matteuccia struthiopteris Dryopteridaceae Underground parts

Onoclea sensibilis* Dryopteridaceae Aerial parts

Onoclea sensibilis* Dryopteridaceae Underground parts

Onoclea sensibilis* Dryopteridaceae Aerial parts

Onoclea sensibilis* Dryopteridaceae Underground parts

Polystichum aculeatum Dryopteridaceae Aerial parts

Polystichum aculeatum Dryopteridaceae Underground parts

Polystichum caryotideum Dryopteridaceae Underground parts

Polystichum falcatum Dryopteridaceae Aerial parts

Polystichum falcatum Dryopteridaceae Underground parts

Polystichum fortunei Dryopteridaceae Aerial parts

Polystichum fortunei Dryopteridaceae Underground parts

Polystichum fortunei Dryopteridaceae Whole plant Polystichum hecatopterum Dryopteridaceae Aerial parts

Polystichum hecatopterum Dryopteridaceae Underground parts

Polystichum lepidocaulon Dryopteridaceae Whole plant Polystichum macrophyllum Dryopteridaceae Underground

parts Polystichum macrophyllum Dryopteridaceae Leaves Polystichum munitum Dryopteridaceae Whole plant + Polystichum polyblepharum Dryopteridaceae Aerial parts

Polystichum polyblepharum Dryopteridaceae Underground parts

Polystichum rigens Dryopteridaceae Whole plant Polystichum setiferum Dryopteridaceae Aerial parts

Polystichum setiferum Dryopteridaceae Underground parts

Polystichum setiferum Dryopteridaceae Whole plant Polystichum tsus-simense Dryopteridaceae Aerial parts

Polystichum tsus-simense Dryopteridaceae Underground parts

Rumohra adiantiformis Dryopteridaceae Whole plant Tectaria decurrens Dryopteridaceae Whole plant Dicranopteris chinensis Gleicheniaceae Whole plant

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Dicranopteris dichotoma Gleicheniaceae Underground parts

Dicranopteris linearis Gleicheniaceae Aerial parts Dicranopteris pectinata Gleicheniaceae Aerial parts Diplopterygium laevissimum Gleicheniaceae Leaves

Diplopterygium laevissimum Gleicheniaceae Underground parts

Diplopterygium laevissimum Gleicheniaceae Stems Sticherus flagellaris Gleicheniaceae Aerial parts Trichomanes hildebrandtii Hymenophyllaceae Whole plant Angiopteris fokiensis Marattiaceae Leaves Angiopteris fokiensis Marattiaceae Stems

Angiopteris fokiensis Marattiaceae Underground parts

Marattia fraxinea Marattiaceae Fronds Marattia fraxinea Marattiaceae Stems Marsilea quadrifolia Marsileaceae Whole plant Marsilea schelpiana Marsileaceae Whole plant Nephrolepis abrupta Nephrolepidaceae Aerial parts

Nephrolepis abrupta Nephrolepidaceae Underground parts

Nephrolepis auriculata Nephrolepidaceae Underground parts

Nephrolepis auriculata Nephrolepidaceae Aerial parts Nephrolepis biserrata Nephrolepidaceae Leaves Nephrolepis biserrata Nephrolepidaceae Stems Nephrolepis biserrata Nephrolepidaceae Fronds Oleandra distenta* Oleandraceae Leaves

Oleandra distenta Oleandraceae Underground parts

Oleandra distenta* Oleandraceae Leaves Oleandra distenta Oleandraceae Stems Ophioglossum thermale Ophioglossaceae Whole plant Osmunda japonica Osmundaceae Leaves

Osmunda japonica Osmundaceae Underground parts +

Osmunda japonica Osmundaceae Stems

Osmunda regalis Osmundaceae Underground parts

Osmunda regalis Osmundaceae Aerial parts

Osmunda vachellii Osmundaceae Underground

parts Osmunda vachellii Osmundaceae Stems Osmunda vachellii Osmundaceae Leaves Plagiogyria adnata Plagiogyriaceae Whole plant Plagiogyria euphlebia Plagiogyriaceae Stems Plagiogyria euphlebia Plagiogyriaceae Leaves Plagiogyria euphlebia Plagiogyriaceae Underground

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

parts

Plagiogyria japonica Plagiogyriaceae Leaves Plagiogyria stenoptera Plagiogyriaceae Underground

parts Plagiogyria stenoptera Plagiogyriaceae Leaves Aglaomorpha coronans Polypodiaceae Whole plant Colysis digitata Polypodiaceae Underground

parts + Colysis digitata Polypodiaceae Aerial parts

Drynaria bonii Polypodiaceae Underground parts

Drynaria fortunei Polypodiaceae Underground parts

Lemmaphyllum microphyllum Polypodiaceae Whole plant Lepisorus asterolepis Polypodiaceae Whole plant Lepisorus contortus Polypodiaceae Whole plant Lepisorus eilophyllus Polypodiaceae Whole plant Lepisorus macrosphaerus Polypodiaceae Aerial parts Lepisorus oligolepidus Polypodiaceae Whole plant Lepisorus tosaensis Polypodiaceae Whole plant Microsorum dilatatum Polypodiaceae Aerial parts +

Microsorum fortuni Polypodiaceae Underground parts

Microsorum fortuni Polypodiaceae Aerial parts Microsorum fortuni Polypodiaceae Whole plant Microsorum membranaceum Polypodiaceae Whole plant

Microsorum pteropus Polypodiaceae Underground parts

Microsorum pteropus Polypodiaceae Aerial parts Microsorum punctatum Polypodiaceae Fronds Neocheiropteris ovatus* Polypodiaceae Whole plant Neocheiropteris ovatus* Polypodiaceae Whole plant Neocheiropteris phyllomanes Polypodiaceae Whole plant Niphidium crassifolium Polypodiaceae Whole plant Platycerium alcicorne Polypodiaceae Leaves Platycerium alcicorne Polypodiaceae Shield leaves Platycerium alcicorne Polypodiaceae Fronds Platycerium alcicorne Polypodiaceae Shield leaves Pleopeltis phymatodes Polypodiaceae Leaves Pleopeltis phymatodes Polypodiaceae Stems

Pleopeltis phymatodes Polypodiaceae Underground parts

Polypodium cambricum Polypodiaceae Whole plant Polypodium decumanum Polypodiaceae Aerial parts

Polypodium decumanum Polypodiaceae Underground parts

Pyrrosia adnascens Polypodiaceae Underground parts

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Pyrrosia adnascens Polypodiaceae Aerial parts Pyrrosia gralla Polypodiaceae Aerial parts + Pyrrosia lanceolata Polypodiaceae Leaves Pyrrosia lingua* Polypodiaceae Whole plant Pyrrosia lingua Polypodiaceae Aerial parts

Pyrrosia lingua Polypodiaceae Underground parts +

Pyrrosia lingua* Polypodiaceae Whole plant Pyrrosia mollis Polypodiaceae Aerial parts Pyrrosia pekinensis Polypodiaceae Aerial parts Pyrrosia petiolosa Polypodiaceae Whole plant Pyrrosia sheareri Polypodiaceae Leaves Acrostichum aureum Pteridaceae Fronds Acrostichum aureum Pteridaceae Stems Adiantum capillus-veneris* Pteridaceae Whole plant Adiantum capillus-veneris* Pteridaceae Whole plant Adiantum caudatum Pteridaceae Whole plant

Adiantum flabellulatum Pteridaceae Underground parts

Adiantum flabellulatum Pteridaceae Aerial parts Adiantum flabellulatum Pteridaceae Whole plant Adiantum raddianum Pteridaceae Aerial parts

Adiantum raddianum Pteridaceae Underground parts

Adiantum venustum Pteridaceae Whole plant Aleuritopteris argentea Pteridaceae Whole plant Cheilanthes duclouxii Pteridaceae Whole plant +

Coniogramme japonica* Pteridaceae Underground parts

Coniogramme japonica* Pteridaceae Underground parts

Coniogramme japonica Pteridaceae Aerial parts Pellaea falcata Pteridaceae Whole plant Pellaea rotundifolia Pteridaceae Whole plant

Pellaea ternifolia Pteridaceae Underground

parts Pellaea ternifolia Pteridaceae Aerial parts Pityrogramma calomelanos Pteridaceae Fronds + Pteris aquilina Pteridaceae Fronds Pteris cretica Pteridaceae Aerial parts Pteris ensiformis Pteridaceae Whole plant

Pteris fauriei Pteridaceae Underground parts

Pteris fauriei Pteridaceae Aerial parts Pteris maclurioides Pteridaceae Whole plant Pteris multifida Pteridaceae Stems

Pteris multifida Pteridaceae Underground parts

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Pteris multifida Pteridaceae Leaves Pteris multifida Pteridaceae Whole plant Pteris multifida Pteridaceae Aerial parts

Pteris multifida Pteridaceae Underground parts

Pteris semipinnata* Pteridaceae Whole plant + Pteris semipinnata* Pteridaceae Whole plant Pteris tremula Pteridaceae Whole plant Anemia phyllitidis Schizaeaceae Whole plant Lygodium flexuosum Schizaeaceae Spores Lygodium japonicum Schizaeaceae Aerial parts Lygodium japonicum Schizaeaceae Whole plant Lygodium kerstenii Schizaeaceae Fronds Lygodium lanceolatum Schizaeaceae Fronds Lygodium scandens Schizaeaceae Aerial parts

Lygodium scandens Schizaeaceae Underground parts

Cyclosorus acuminatus Thelypteridaceae Whole plant

Cyclosorus dentatus Thelypteridaceae Underground parts

Cyclosorus dentatus Thelypteridaceae Aerial parts

Cyclosorus falcilobus Thelypteridaceae Underground parts +

Cyclosorus falcilobus Thelypteridaceae Stems + Cyclosorus falcilobus Thelypteridaceae Fronds + Cyclosorus interruptus Thelypteridaceae Fronds

Cyclosorus interruptus Thelypteridaceae Underground parts

Cyclosorus parasiticus Thelypteridaceae Whole plant + Cyclosorus penangianus Thelypteridaceae Whole plant

Cyclosorus tylodes Thelypteridaceae Underground parts

Cyclosorus tylodes Thelypteridaceae Aerial parts

Phegopteris decursive-pinnata Thelypteridaceae Underground parts +

Phegopteris decursive-pinnata Thelypteridaceae Leaves Thelypteris beddomei Thelypteridaceae Leaves Thelypteris beddomei Thelypteridaceae Stems + rhizome Thelypteris glanduligera* Thelypteridaceae Whole plant + Thelypteris glanduligera* Thelypteridaceae Whole plant Vittaria isoetifolia Vittariaceae Whole plant + *Samples in duplicate refer to different collections.

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Table 2S Major phenotypic changes induced by 13 reference compounds including anticancer

drugs or mitotic blockers

The compounds were chosen for their various mechanisms of action to validate the phenotypic

assay. They were evaluated at two concentrations corresponding to 2 x and 4 x IC50. First

observations of the global population were performed including the evaluation of cell density,

proportion of mitotic cells and cell death. Then effects on interphasic and mitotic cells labelled for

-tubulin, -tubulin and DNA were evaluated.

In the column “phases”, the underlined term corresponds to the predominant phase in the mitotic

population.

Topo=topoisomerase, PM=prometaphase, M=metaphase, Lc= lagging chromosomes

(1) Hastie SB. Interactions of colchicine with tubulin. Pharmacol Ther 1991; 51: 377-401 (2) Kruczynski A, Barret JM, Etievant C, Colpaert F, Fahy J, Hill BT. Antimitotic and

tubulin-interacting properties of vinflunine, a novel fluorinated Vinca alkaloid. Biochem Pharmacol 1998; 55: 635-648

(3) Ootsu K, Kozai Y, Takeuchi M, Ikeyama S, Igarashi K, Tsukamoto K, Sugino Y, Tashira T, Tsukagoshi S, Sakurai Y. Effects of new antimitotic antibiotics, ansamitocins, on the growth of murine tumors in vivo and on the assembly of microtubules in vitro. Cancer Res 1980; 40: 1707-1717

(4) Samson F, Donoso JA, Heller-Bettinger I, Watson D, Himes RH. Nocodazole action on tubulin assembly, axonal ultrastructure and fast axoplasmic transport. J Pharmacol Exp Ther 1979; 208: 411-417

(5) Schiff PB, Fant J, Horwitz SB. Promotion of microtubule assembly in vitro by taxol. Nature 1979; 7: 665-667

(6) Mayer TU, Kapoor TM, Haggarty SJ, King RW, Schreiber SL, Mitchison TJ. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 1999; 286: 971-974

(7) Ross W, Rowe T, Glisson B, Yalowich J, Liu L. Role of topoisomerase II in mediating epipodophyllotoxin-induced DNA cleavage. Cancer Res 1984; 44: 5857-5860

(8) Hsiang YH, Liu LF. Identification of mammalian DNA topoisomerase I as an intracellular target of the anticancer drug camptothecin. Cancer Res 1988; 48: 1722-1726

(9) Aubel-Sadron G, Londos-Gagliardi D. Daunorubicin and doxorubicin, anthracycline antibiotics, a physicochemical and biological review. Biochimie 1984; 66: 333-352

(10) Hoshikawa Y, Kwon HJ, Yoshida M, Horinouchi S, Beppu T. Trichostatin A induces morphological changes and gelsolin expression by inhibiting histone deacetylase in human carcinoma cell lines. Exp Cell Res 1994; 214: 189-197

(11) Kovacs M, Toth J, Hetenyi C, Malnasi-Csizmadia A, Sellers JR. Mechanism of blebbistatin inhibition of myosin II. J Biol Chem 2004; 279: 35557-35563

(12) Harrington EA, Bebbington D, Moore J, Rasmussen RK, Ajose-Adeogun AO, Nakayama T, Graham JA, Demur C, Hercend T, Diu-Hercend A, Su M, Golec JM, Miller KM. VX-680, a potent and selective small-molecule inhibitor of the Aurora kinases, suppresses tumor growth in vivo. Nat Med 2004; 10: 262-267

(13) Adams J, Behnke M, Chen S, Cruickshank AA, Dick LR, Grenier L, Klunder JM, Ma YT, Plamondon L, Stein RL. Potent and selective inhibitors of the proteasome: dipeptidyl boronic acids. Bioorg Med Chem Lett 1998; 8: 333-338

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© Georg Thieme Verlag KG · DOI 10.1055/s-0030-1250407 · Planta Med · Pouny I et al.

Cell density Mitotic cells Cell death Cell morphology nuclei -tubulin centrosomes Phases chromosomes mitotic spindle centrosomes

x 2 low low blebbing irregular-shaped nuclei + disorganized microtubule network normal or surnumerary PM, M Lc in M 2 close poles of microtubules or

more than 2 polesnormal or

supernumerary

x 4 very low high blebbing irregular-shaped nuclei + disorganized microtubule network normal or surnumerary PM normal no microtubule normal

x 2 low low rounded cells ++ irregular-shaped nuclei ++; micronuclei ++

disorganized microtubule network normal or surnumerary PM, M Lc in M close poles or 3 poles supernumerary

x 4 very low low rounded cells + fragmented nuclei; micronuclei ++

disorganized microtubule network surnumerary PM normal multipolar. Low density of

microtubules. supernumerary

x 2 high = very low rounded cells + irregular-shaped nuclei + normal normal or surnumerary all Lc in M close poles supernumerary

x 4 medium low rounded cells ++; protrusions

irregular-shaped nuclei +; fragmented nuclei ++

microtubules in protrusions and cytoplasmic bridges normal or surnumerary PM, M Lc in M close poles or 3 poles supernumerary

x 2 medium low normal fragmented nuclei; micronuclei + disorganized microtubule network surnumerary PM normal multipolar supernumerary

x 4 medium low normal fragmented nuclei; micronuclei + disorganized microtubule network surnumerary PM normal multipolar supernumerary

x 2 medium low normal fragmented nuclei; micronuclei + normal surnumerary and scattered all, PM Lc in M multipolar normal or supernumerary

x 4 very low low rounded cells + fragmented nuclei; micronuclei + disorganized microtubule network surnumerary and scattered PM normal multipolar, spindles of dense

microtubules supernumerary

x 2 low very low rounded cells + irregular-shaped nuclei + disorganized microtubule network normal P, PM normal monopolar gathered

x 4 very low high rounded cells ++ irregular-shaped nuclei ++; fragmented nuclei +

disorganized microtubule network normal PM hypercondensed

chromosomes monopolar gathered

x 2 low none low normal binucleation normal 2 scattered centrosomes or surnumerary

x 4 very low none high rounded cells ++ low density nucleimicrotubule ring around nuclei,

disorganized microtubule network

3 scattered centrosomes or surnumerary

x 2 low none very low normal micronuclei + microtubule ring around nuclei normal

x 4 low none low rounded cells + binucleation; micronuclei + microtubule ring around nuclei normal

x 2 medium none very low rounded cells + binucleation normal normal

x 4 medium none very low rounded cells ++; protrusions; blebbing micronuclei + microtubule ring around nuclei normal

x 2 medium = very low protrusions + normal 1 to 3 protrusions with microtubules per cell normal all Lc in M mainly normal spindles. Some

tripolar PM normal

x 4 low low protrusions ++ irregular-shaped nuclei + 1 to 4 protrusions with microtubules per cell normal PM normal multipolar supernumerary

x 2 very high = low protrusions, cytoplasmic bridges binucleation microtubules in protrusions and

cytoplasmic bridgessurnumerary in binucleated

cells all normal wide midbody normal

x 4 very high = high protrusions, cytoplasmic bridges binucleation microtubules in protrusions and

cytoplasmic bridgessurnumerary in binucleated

cells all normal wide midbody normal

x 2 high = very low blebbing fragmented nuclei +; polylobed nuclei +

disorganized microtubule network surnumerary PM normal monopolar normal

x 4 high low blebbing fragmented nuclei +; polylobed nuclei +

disorganized microtubule network surnumerary PM normal monopolar; short microtubules normal

x 2 high very low rounded cells ++; protrusions normal disorganized microtubule

network surnumerary PM, M normal monopolar normal

x 4 high none low rounded cells ++; protrusions normal disorganized microtubule

network none or surnumerary none

Interphase

Camptothecin (1,4.10-6 M)

Doxorubicin (2,0.10-7 M)

Trichostatin A (8,0.10-8 M)

Ansamitocin P-3 (5,7.10-11 M)

Vinflunine (3,4.10-8 M)

Taxol (2,0.10-9 M)

Monastrol (5,1.10-5 M)

HDACs (10)

Kinesin Eg5 (6)

tubulin (2)

tubulin (3)

tubulin (5)

Blebbistatin (>1,0.10-5 M)

Myosin (11)

Topo. II (7)

Topo. I (8)

DNA (9)

Nocodazole (4,7.10-8 M)

tubulin (4)

Colchicine (7,0.10-9 M)

General qualitative observationsTreatment (IC50 on

HeLa cells)IC50

ratioTarget

tubulin (1)

PS341 (9,8.10-8 M)

Proteasome (13)

Etoposide (3,2.10-5 M)

VX680 (4,2.10-7 M)

Aurora B (12)

none

Mitosis

none

none